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‘Warrior’ (Reg. No. CV-252, PI 655523), ‘Scout’ (Reg. No. CV-253, PI 655524), and ‘Chief’ (Reg. No. CV-254, PI 655525) indiangrass [Sorghastrum nutans (L.)] Nash were developed for improved yield and forage digestibility by USDA-ARS and the University of Nebraska for use in the central Great Plains and the Midwest USA. Warrior was tested as Oto C3 and was developed by means of both half-sib family selection and restricted, recurrent phenotypic selection (RRPS). Scout and Chief which were tested as NE54 C2 and Holt × Oto Early C2, respectively, were developed by RRPS. Warrior is adapted to USDA Plant Hardiness Zone (HZ) 5 and the upper part of HZ 6 in the Great Plains and Midwest. It produces forage with high in vitro dry matter digestibility (IVDMD) that results in improved animal gains when utilized by beef cattle in well managed grazing systems in regions where it is adapted. Scout is adapted to HZ 5 in the Great Plains and Midwest, USA. It produces signifi cantly greater forage yields than other adapted indiangrass cultivars when grown for hay in the western part of its adaptation region. Chief is adapted to HZ 4 and the upper half of HZ 5. It produces signifi cantly greater forage yields than the other available HZ 4 cultivars. Warrior, Scout, and Chief represent the second generation of indiangrass cultivars developed for use in production agriculture. In the regions where they are adapted, these cultivars can be used in pure stands or in multispecies mixtures with other grasses.

Grain sorghum [Soirghum bocolor (L.) Moench] typically produces larger seeds than sudangrass [S. bicolor, formerly S. sudanense (Piper) Stapf]; and grain sorghum seedlings are higher in hydricyanic acid potential (HCN-p) than sudangrass seedlings. Previous studies have shown a seed-parent effect on seed weight and HCN-p in reciprocal F 1 hybrids of sorghum × sudangrass

Spectrophotometry is a useful assay for hydrocyanic acid potential (HCN-p) in sorghum and sudangrass [both Sorghum bicolor (L.) Moench] seedlings, but no systematic study of sources of variation in the procedure has been reported. Selfed seed was harvested from each of 12 ramets (two each from two sister plants from three low-HCN-p sudangrass parents), and seedlings for rumpling were grown in a growth chamber, in two rows from each ramet. Seven-day-old seedlings were harvested and divided into two samples per row for extraction; two aliquots per extract were assayed spectrophotometrically for HCN-p. The experiment was replicated three times. The three parents differed in HCN-p [...]

Two field studies involving different groups of germplasm were conducted to compare sorghum-sudangrass, Sorghum bicolor (L.) Moench X S. sudanense (Piper) Stapf, hybrid seed production of male-sterile FI's with their component A·lines. The FI's yielded 45 and 82% more grain than their A·line counterparts in Study I and 2, respectively, with the increase attributable to more seeds per panicle and a greater threshing percentage. Mean values of nine traits measured on nine groups of eight FI's in Study I and seven groups of six FI's in Study 2 were compared with their respective A·lines. All groups of FI's significantly outyielded their A·line counterparts in both studies except for A·line N35 in Study 1. Phenotypic correlations of yield and seeds per panicle were high in both A·lines and FI's in both studies, as were the correlations of yield and threshing percentage except for the FI's in Study 1. Application of these findings should facilitate the production of higher yields of sorghum-sudangrass hybrid seed at reduced cost per unit of seed. The best FI's to use and the magnitude of their superiority over A·lines will be influenced by the location in which seed is produced. Additional indexwords:Sorghum bicolor (L.) Moench, Sorghum sudanense (Piper) Stapf, Single-cross hybrids, Three-way hybrids. SORGHUM-SUDANGRASS, Sorghum bicolor (L.) Moench X S. sudanense (Piper) Stapf, hybrids are grown extensively to provide supplementary forage for animals as pasture, silage, or greenchop. These hybrids play an important role in the management plans of many livestock producers, particularly in drouth-prone regions such as the Great Plains. In a 1977 survey by Harvey (I), 81.5% of the commercially produced sorghum-sudangrass hybrids in the United States were F I 's, with 'Redlan' grain sorghum being the preferred male-sterile and 'Greenleaf sudangrass the most often used pollinator. Also, three-way cross hybrids made up I I% of the total seed production of sorghums and sudangrasses used for forage compared with less than 2% in grain sorghum (I). Despite this rather extensive use of three-way crosses in forage and sudangrass hybrids, only two preliminary reports were found in which the seed production of three-way and single cross hybrids was compared. In a I-year study of seed production of sorghum-sudangrass hybrids in Hungary (3), seed yields of F I male steriles were greater and production was more dependable than for Alines because the F1's matured earlier, but comparative seed yields of three-way and single cross hybrids were not presented. In Japan (6), four grain sorghum A-lines, six grain-sorghum male-sterile F I 's and six sorgo male-sterile F I 's were used in a comparative I Contribution of USDA-ARS and the NebraskaAgric. Exp. Stn. Published asPaper no. 7415, Journal Series, Nebraska Agric. Exp, Stn., Lincoln. Research was conducted under Nebraska Project no. 12-114. Received 29 Feb. 1984. 2 Supervisory research geneticist, USDA-ARS; research technologist inagronomy;George Holmes professorof agronomy; and research geneticist, USDA-ARS, Lincoln, NE68583, respectively. study of seed production of forage sorghum hybrids. Seed weight per head on the grain-sorghum malesterile F I 's was slightly lower than for the A-lines, while seed weight per head on the sorgo male-sterile F I 's was substantially greater than on either of the other two female types. Seed yield per land area was not given. The A-lines used differed in each of the three groups of material, and seed weights of F I 's could not be compared with those of their component A-lines. Although published information on comparisons of three-way with single cross forage-type hybrids is limited, more extensive literature is available for grain sorghums as indicated in a recent report (2). The objectives of this study were to I) compare seed yield and other agronomIC traits between malesterile F I seed parents and their component A-line seed parents in the production of sorghum-sudangrass hybrids in two different groups of germplasm, 2) determine phenotypic correlations among traits within parental groups, and 3) relate the findings to hybrid seed production. Evaluation of the hybrid seed produced in terms of forage production will be reported in a subsequent paper. MA TERIALS AND METHODS The seed parents used in Study 1 were nine combineheight cytoplasmic male-sterile A-lines and their 36 malesterile AX B-line crosses (FI'S) that were identical to those used in a similar study with grain sorghum hybrids (2). The A-lines included KS4, KS23, 'Martin', N30, N35, N36, N38, WD4, and 'Wheatland'. In Study 2, the seed parents were seven A-lines and their 21 male-sterile F t's. The A-lines were KS5 and KS9, described by Ross et al. (4); N38 and N48, described by Ross et al. (5); N4692, described by Webster et al. (7); N50I3, an experimental line; and the cultivar Redlan. The pollen source was produced from a composite of equal seed weights of the sudangrass cultivars Greenleaf and 'Piper', Nebraska 7035, and experimentallow-dhurrin strains of Greenleaf and Piper. This heterogeneous pollen source assured continuous and adequate pollen dispersal to the female parents over the range of their stigma receptivity. The sudangrass composite was seeded in double rows, with six rows of male-sterile sorghums being grouped between each pair of sudangrass rows. The outside rows of each group of six male-steriles were seeded to cytoplasmic male-sterile 'Combine Kafir-60', which served as unharvested border rows to minimize any effect of the taller sudangrass rows on the four randomly assigned male-sterile lines and F I hybrids that were harvested in each group. The experiments were planted 29 May 1979 and 27 May 1980 at the University of Nebraska Field Laboratory, Mead, in a medium textured Sharpsburg silty clay loam soil (fine, montmorillonitic, mesic Typic Argiudoll) to which 112 kg haI of N had been applied. Four replications of singlerow plots 7.6 m long and 0.76 m apart in a randomized complete block design were used each year. All plots were overseeded and thinned to 15 em between plants, giving a plant population of about 87 000 plants ha1• The exper1134 Published in Crop Science (November-December 1984) v.24, no. 6

A method of assessing the hydrocyanic acid potential (HCN-p) of sudangrass [Sorghum sudanense (Piper) Stapf] and sorghum [S. bicolor (L.) Moench] seedlings is described. This procedure is based on the determination of p-hydroxybenzaldehyde (p-HB), which is released upon hydrolysis of dhurrin, the cyanogen normally present in plants of Sorghum species. Extraction and hydrolysis of dhurrin are accomplished by autoclaving young leaf tissue in water. The content of p-HB in the aqueous extract is then determined by spectrophotometric assay in alkaline solution at 330 nm. Uniform samples for the comparison of widely divergent genotypes are obtained by using the first leaf of young, chamber-grown, green seedlings. Relative ranks of a wide variety of sorghum germplasm assayed for HCN content by this technique are in good agreement with those obtained by other methods. The procedure shows promise of providing a rapid and precise tool for conducting genetic and breeding studies with sorghums

The variation among 49 F1 forage sorghum [Sorghum bicolor (L.) Moench] hybrids from a 7 x 7 cross-classified design was explored in 1979 and 1980 for the following traits: dry matter (DM), crude protein (CP), in vitro dry matter disappearance (IVDMD), neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). Differences were found among hybrids for all traits. Parental means for males generally had a wider range of variation than for females. The means of hybrids involving the male parent, 'White Collier', were highest in IVDMD, and lowest in NDF, ADF, and ADL. IVDMD was the only trait that showed significant differences among hybrids averaged over both males and females. General combining ability (GCA) effects were more important than specific combining ability effects. Our results suggest that the most rapid way to improve forage sorghum quality would be by improving IVDMD. Significant differences among hybrids averaged over both male and female parents were shown for this trait; it was affected relatively little by year effects and exhibited high GCA effects.

In a previous report the progress made in altering the in vitro dry matter digestibility (lVDMD) of switchgrass (Panicum oir­ gatum L.) in one cycle of divergent selection was described. In this study, the detergent system of analyses was used in an at­ tempt to determine what plant constituents had been altered by the selection for IVDMD. Hays from five switchgrass strains differing in IVDMD harvested over a 3-year period were ana­ lyzed for protein content, neutral detergent fiber (NDF), acid detergent fiber (ADF), ash, and lignin. Although there were dif­ ferences (p :s 0.05) among strains for IVDMD as determined by two different procedures (with and without additional N), there were no differences among strains for ADF, ash, lignin, and protein. The strains ranked the same with the two IVDMD procedures but the IVDMD·N (with additional N) test averaged 6.7 percentage points higher in digestibility. There were small differences (p -s 0.1) among strains for NDF in 2 of the 3 years. The strain with the highest IVDMD had the lowest NDF, sug­ gesting that, in part, the improved IVDMD was due to a decrease in the cell wall component of the harvested forage. Hays ofthree of these strains, high-IVDMD PC, low-IVDMD PC, and 'Path· finder' from the 1980 harvest were ground and fed to sheep to compare in vivo and in vitro digestibilities. After grinding, the hayshad IVDMD-N values of49.9,45.5, and 46.7%, respectively. There were no differences among strains in the sheep feeding trial for in vivo digestibility, dry matter intake, and fiber di­ gestibility. The in vivo digestibilities for these strains were: high­ IVDMDPC 50.4%, low-IVDMD PC 49.0%, and Pathfinder 50.8%. In contrast to a previous report in the literature, the IVDMD­ N values were generally similar to in vivo digestibilities for switchgrass hays. Parameters of the conventional detergent sys­ tem of analyses, while useful in identifying factors responsible for changes in digestibility of switchgrass hays, would probably be of little value as selection criteria in a breeding program. Because of the large number of animals required to obtain sta­ tistical significance in in vivo digestibility studies, the IVDMD procedure would be preferable in studies in which breeding ma­ terial is being evaluated and small differences in digestibility are expected.

Alteration in hydrocyanic acid potential (HCN-p) of the forage is one of the important objectives in sudangrass [Sorghum sudanense (Piper) Stapf] breeding programs. The effectiveness of recurrent phenotypic selection for increasing or decreasing HCN-p in sudangrass was evaluated in two cycles of individual plant selection in the cultivar 'Greenleaf'. In cycle 1, HCNP means of the high and low populations were higher and lower, respectively, than for Greenleaf, but only the low-HCN-p population was significantly different from Greenleaf. In cycle 2, mean HCN-p values of both populations differed significantly from Greenleaf. The average realized heritability for the two cycles was 0.40 while broad-sense heritability estimates averaged 0.86. After two cycles of selection, the low and highHCN-p populations differed from Greenleaf by about 17 and 30%, respectively. Additional index words: Prussic acid, Dhurrin, Forage quality, Heritability, Sorghum sudanense. SU D A N G R A S S [Sorghum sudanense (Piper) Stapf] and sorghum-sudangrass hybrids are used extensively to provide supplementary feed to animals as pasture or greenchop. Precautions in managing the crop are necessary to prevent animal losses due to prussic acid (hydrocyanic acid) poisoning. All known sudangrasses and sorghums [SO bicolor (L.) Moench] contain dhurrin [(S)-phydroxymandelonitrile (J-D-glucopyranoside] which yields hydrocyanic acid when hydrolyzed enzymatically in disrupted plant tissues or in the rumen of consuming anim';lls. Breed.ing sudangrasses with lowered hydrocyanic acid potential (HCN-p) would reduce the danger of hydrocyanic acid poisoning, and permit greater flexibility in the management of this crop. Also, sudangrasses with reduced HCN-p would be useful in the development of sorghum-sudangrass hybrids with lowered HCN-p. 'Contribution from USDA-ARS and the Nebraska Agric. Exp. Stn., ~incoln. Published as Paper No. 6173, Journal Series, Nebraska Agric. Exp. Stn. Received 18 Feb. 1981. The work reported was conducted under Nebraska Agric. Exp. Stn. Projects 12-088 and 12-114. 'Supervisory research geneticist, USDA-ARS; George Holmes professor of Agronomy, Univ. of Nebraska; and research agronomist, USDA-ARS, Lincoln, NE 68583, respectively. GORZ ET AL.: HYDROCYANIC ACID POTENTIAL IN SUDANGRASS 323 Breeding for lower HCN-p in sudangrass was facilitated by the recent development of a simple, rapid, nondestructive spectrophotometric procedure (6). This procedure involves the assay of individual first leaves from 7-day-old seedlings grown under controlled conditions. Following completion of the assay, selected seedlings can be transplanted to the field or greenhouse for the production of self or cross-pollinated seed for use in the next cycle of selection. In a review of studies of the inheritance of cyanogenesis in sudangrass and sorghum, Nass (9) reported that dominant or partially dominant factors were involved in the genetic control of both high and low HCN-p. Most studies suggested multigenic inheritance although one or two major genes also were hypothesized. Hogg and Ahlgren (8) evaluated 175 inbred lines ranging from low to high in HCN content during a 3-year period and reported that HCN content of the lines was stable over years. They also reported that low-HCN strains could be developed by crossing low-HCN inbred lines. Barnett and Caviness (1) reported broad-sense heritability estimates of 0.41 and 0.68 for HCN production in populations derived from two sorghum X sudangrass crosses. Sorghum is classified as a predominately self-pollinated crop with outcrossing averaging 6% (10). The outcrossing percentage of sudangrass is usually higher, but reported values vary quite widely. Garber and Atwood (5) observed 76.4, 18.2, and 34.4% cross-pollination in Pennsylvania for the years 1941, 1942, and 1943, respectively, while Hogg and Ahlgren (8) reported in 1943 that cross-pollination in Wisconsin ranged from 4.5 to 10%. Thus, extensive cross-pollination is possible, but the percentage that occurs in a specific seed field is apparently dependent upon the location, environmental factors, and the type of sudangrass being grown. Recent work by Foster et al. (4) demonstrated positive results for bi-directional mass selection in a grain sorghum population. Thus, selection based on population improvement techniques, developed for use with cross-pollinated crops, may be successfully used with crops such as sorghum that have a very low percentage of cross pollination. 'Greenleaf sudangrass is used extensively as the male parent in commercial production of sorghum-sudangrass hybrids (7). In 1977, a program of recurrent selection for low and high HCN-p in Greenleaf sudangrass was initiated at the Nebraska Station. The objectives of this research were to evaluate the effectiveness of recurrent selection for increasing or decreasing HCN-p in sudangrass populations, and to develop a reselected Greenleaf strain with lower HCN-p than the parent. MATERIALS AND METHODS All HCN-p values in this study were obtained by use of the spectrophotometric procedure (6). In this procedure, first-leaf samples from week-old seedlings were weighed, and dhurrin was extracted and hydrolyzed by autoclaving the samples in water. Aliquots of the extracts were then diluted in base, and absorbance was read at 330 nm, the absorption maximum of p-hydroxybenzaldehyde. HCN-p values were derived from the 330 nm absorbance (A330) readings by simple calculations (6). The initial population consisted of 184 seedlings of the cultivar Greenleaf grown from Kansas foundation seed. From this initial population, 19, 17, and 17 seedlings were selected to represent the lowest, intermediate, and highest HCN-p levels in further studies. The selected seedlings were transplanted to the field, heads were bagged prior to anthesis, and selfed seed was harvested. Five plants from each HCN-p group with adequate supplies of selfed seed were selected for further study. Seven replications of the seed from the 15 selected plants were planted for assay of HCN-po A total of 1405 S, seedlings were assayed with a range of 81 to 110 per line. This phase of the study will be referred to as cycle O. For the next step, cycle 1, only those seedlings with the highest and lowest levels of HCN-p were selected from the total of 1405 S, seedlings obtained from the 15 original plants. Selections were made across all replications and without regard to the original HCN-p group (high, medium, or low) from which the seedling originated. Thus, the 26 seedlings selected for lowHCN-p included 14, 9, and 3 seedlings from the low, medium, and high-HCN-p groups, respectively. Similarly, the 23 seedlings selected for high-HCN-p included 9, 5, and 9 seedlings from the low, medium, and high-HCN-p groups, respectively. The low and high-HCN-p populations were transplanted to separate field isolations in 1978, with plants on 61-cm centers. Open-pollinated seed was harvested from individual plants. Three replications of 10 seedlings each from each line (i.e., from each parent plant) were assayed for HCN-p. Greenleaf was included in each replication as a control. Plants for the next cycle of selection (cycle 2) were selected from individual seedlings of the 8 lines whose mean HCN-p was lowest among the 26 low-HCN-p lines, and from the highest 8 lines among the 23 high-HCN-p lines. The selected low and high-HCN-p populations included 54 and 56 plants, respectively. Each population was transplanted to a separate field isolation in 1979, with plants randomized on 107-cm centers. A severe infestation of chinch bugs [Blissus leucopterus leucopterus (Say)] destroyed some plants in the low-HCN-p isolation; seed was produced on only 33 of these plants. Open-pollinated seed was harvested from individual plants, and three replications of 10 seedlings from each plant that produced sufficient seed (31 plants from the low-HCN-p isolation and 55 from the highHCN-p isolation) were assayed for HCN-p. Greenleaf was included in each replication as a check. The breeding procedures used and the time period covered by each cycle are summarized in Table 1. A narrow-sense heritability estimate was obtained by the regression of offspring produced by selfing on their parents in cycle O. In this situation, bop, the regression of offspring on parents, was used as the heritability estimate (H,,). Realized heritability estimates (Hr) (3) were also determined for gcles 0, 1, ~d 2 using thefollowing equation: Hr = CX.,h Xo,)/(Xp" X p,) where the X's are means and the subscripts, 0, p, h, and 1 are offspring, parent, high and low, respectively. Variance components from the analyses of the replicated tests of the progeny of each cycle were used to calculate the ratio: Hv = a a'e)where a a the genetic and environmental components of variance, respectively. This ratio was difficult to interpret for this study because it was unknown whether the parents were homozygous or heterozygous individuals or whether the progenies were the products of crossing, selfing, or both. It is an estimate, however, of the variance among lines that is due to genotypic differences and as such can be used as a broad-sense heritability estimate. Gain from selection (Gs) was calculated both as a deviation from Greenleaf (G",) and as a deviation percentage from Greenleaf (G,p) for cycles 1 and 2. The equations used were as follows: (a) G'd for low HCN-p = (X-J X~. (b) G'd for high HCN-p = QCOh -_XgL (c) G,p for low HCN-p = Ph X~/~ X 100. (d) G,p for high HCN-p = (X"h Xg)/Xg X 100. 324 CROP SCIENCE, VOL. 22, MARCH-ApRIL 1982 Table 1. Means, standard deviations, and ranges for HCN·p of parents and their progenies for two cycles of divergent selection for HCN·p in Greenleaf sudangrass.

The hydrocyanic acid potential (HCN-p) of sorghum [Sorghum bicolor (L.) Moench] plants is recognized as a heritable trait, but previous tudies on the mode of inheritance of HCN-p have produced inconsistent results. The objective of this study was to investigate the inheritance patterns of seedling HCN-p and also of seed weight in reciprocal crosses of sorghum and sudangrass [formerly S. sudanense (Piper) Stapf]. Both traits were found to be inherited quantitatively. Generation means analysis indicated that additive genetic effects were most important for both seed weight (53% of variation) and seedling HCN-p (74% of variation). A maternal effect was found for both traits in the F~ and backcross generations. No evidence of this reciprocal effect was found in the F2, suggesting that cytoplasmic inheritance was not involved. A highly positive correlation between seed weight and HCN-p was found across all entries, across both parents, and in the F~ and backcross generations. However, correlation coefficients between seed weight and seedling HCN-p for individual entries, the pooled F2’S, or within types of seed parents in the F~ or backcross generations were generally nonsignificant. It was concluded that seed weight per se does not have a large effect on seedling HCN-p. Additional index words: Dhurrin, Prussic acid, Sorghum blcolor (L.) Moench, S. sudanense (Piper) Stapf. p REVIOUS studies of the inheritance of the cyanogenic glucoside, dhurrin [p-hydroxy-(S)-mandelonitrile-#-D-glucoside], in sorghum [Sorghum bicolor (L.) Moench] have not yielded consistent results. Nass (17) in 1972 reviewed published reports on the inhero itance of cyanogenesis in sorghum, various Lotus species, and white clover (Trifolium repens L.), and concluded that the situation in sorghum was more complex than that in other species. The studies agreed that the dhurrin content or hydrocyanic acid potential (HCNp) of sorghum leaves was a heritable trait, but disagreed on the presence or absence of dominance, dominance of high or low HCN-p, and the number of genes involved. Krauss (14), in a study not included in Nass’s review, concluded that HCN-p was conditioned by four genes with additive effects without dominance. Other studies conducted since Nass’s review reported different conclusions on the importance of additive and nonadditive effects (2, 4, 9). Recently, Gorz et al. (10) found that seedlings of sorghum lines KS8 and N32 were high in HCN-p, but the HCN-p of flag leaves from field-grown plants of KS8 was only about 1/10 as high as that of N32 flag leaves. A single gene pair was found to be primarily responsible for the large difference in mature leaves, and neither low nor high HCN-p was completely dominant. The lack of agree~ Contribution from the USDA-ARS and the Nebraska Agric. Res. Div., Lincoln, NE 68583. Published as Paper no. 8119, Journal Series, Nebraska Agaric. Res. Div. Research was conducted under Project 12-114. Recetved 7 Aug. 1986. -~ Former graduate research assistant, George Holmes professor of agronomy, and supervisory research geneticists, USDA-ARS, respectively. Published in Crop Sci. 27:522-525 (1987). ment among these studies was no doubt due in part to differences in lines or cultivars used, conditions of growth, types of tissue assayed, and the analytical procedures used. The primary objective of the present study was to investigate the inheritance of seedling HCN-p in crosses of grain sorghum and sudangrass [formerly S. sudanense (Piper) Stapf] lines. The sudangrass lines had been selected for low HCN-p; crosses of these lines to the high-HCN-p sorghum lines provided a wider range in HCN-p than was possible in most of the previous studies of HCN-p inheritance. Also, this study utilized the spectrophotometric assay described by Gorz et al. (8). This assay is independent of the activity of catabolic enzymes in the plant. MATERIALS AND METHODS Four inbred parental lines were used in this study. Two were lines from sudangrass populations, and two were grain sorghum cultivars. The two sudangrass lines, 81-1901-7 and 81-1904-74, hereafter referred to as 1901 and 1904, respectively, were selected primarily for low HCN-p. Both lines carried the ms3 gene for genetic male sterility (gms); this recessive gene caused 25 to 30% of the plants in each line to be sterile. Both sudangrass lines also restored fertility when crossed with cytoplasmic-male-sterile (cms) sorghum lines having A1 cytoplasm. The population from which line 1901 was selected has since been released and registered as NP25 low-dhurrin sudangrass germplasm (12). Line 1904 was selected from a population with most of its background from ’Piper’. Both 1901 and 1904 are very low in seedling HCNp compared to commercially available sudangrasses, and they are considered to be homozygous for low HCN-p. The two grain sorghums used were ’Redlan’ and ’Combine Kafir 60’ (CK60). A-lines (A 1 cytoplasm) and B-lines of these cultivars have been maintained for at least 10 yr by making paired crosses annually to produce A-line seed and also selfing the B-line plants used in the A-line crosses. The lines are considered to be homozygous for high seedling HCN-p. All possible crosses, including reciprocals, were made among the four parental lines. Crosses were made using gms plants from the sudangrass lines and cms plants from the sorghum cultivars as females. Use of these male-sterile plants allowed for production of a large number of FI seeds with minimal risk of contamination during crossing. The FI plants were selfed to produce F2 seed, and were also used as pollen sources in backcrosses to gms sudangrass and cms sorghum parental plants. To get backcross seed from the sorghum × sorghum F~’s, the cms hybrids were used as females, and pollen was taken from the parental sorghum B-lines. All crosses were made at the University of Nebraska Agronomy Farm, Lincoln, NE, in the summers of 1982 and 1983 or in the greenhouse in early 1984. All seedlings analyzed in the laboratory were grown from seed produced either in the field in 1983 or in the greenhouse in early 1984. In total, 52 entries were assayed in the laboratory, including 12 F~’s, 10 F2’s, 24 backcrosses, and 6 parental lines (1901, 1904, ARedlan, BRedlan, ACK60, and BCK60). Twenty-five seed packets were prepared for each of the 52

Vanillin-HCl procedures are widely used for the assay of tannins in plants. In attempts to adapt such procedures for use with sorghum [Sorghum bicolor (L.) Moench.] forage it was found that false positive reactions resulted, that is, red color developed in the presence of HCl with or without vanillin. Leucoanthocyanidins (monomeric proanthocyanidins) may be the constituents responsible for this red color. A "chloroform-HCl" procedure was developed for measuring leucoanthocyanidins in sorghum forage. The procedure avoids interference by chlorophyll or other chloroform-soluble constituents. With vanillin added to the solution, this procedure should also be useful for the assay of condensed proanthocyanidins (tannins). Additional index words: Sorghum bicolor L. Moench., Leucoanthocyanidin, Proanthocyanidin, Forage quality. A indicated by Sarkar et al. (14), tannins may be regarded as desirable (for their possible protection against bloat in grazing animals) or undesirable (for their adverse effect on digestibility) constituents of forages. Therefore, either increased or decreased tannin content might be a reasonable breeding objective, depending upon the nature and proposed use of the crop in question. Whether working toward increased or reduced levels of tannins, the breeder needs a simple, rapid, and reliable procedure for tannin assay. Vanillin-HCI procedures such as that described by Burns (2) have been developed for this purpose. In this procedure, dried and ground plant samples are extracted with methanol, and portions of the resulting extract are reacted with a solution containing vanillin and HCI in methanol. The development of a red color indicates the presence of tannins. Vanillin-HCI and vanillin-H2S04 reagents have been used extensively for the detection and assay of tannins and related substances in leguminous forage crops (2, 8, 13, 14) and in sorghum [Sorghum bieolor (L.) Moench] grain (1, 3,5,9, 11, 12). Reports of the use of vanillin-HCI proI Contribution from ARS, USDA, and the Dep. of Agronomy, Nebraska Agric. Exp. Stn., Lincoln. Published as Paper No. 6680, Journal Series, Nebraska Agric. Exp. Stn. Received 22 Apr. 1982. The work reported was done under Nebraska Agric. Exp. Stn. Project 12-114. 2 Former graduate research assistant; George Holmes professor of agronomy; and supervisory research geneticist, ARS-USDA, and professor of agronomy, respectively; University of Nebraska, Lincoln, NE 685830915. Current address of the senior author: FFR Cooperative, West Lafayette, IN 47906. 3 Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by USDA or the Univ. of Nebraska, and does not imply its approval to the exclusion of other products that may also be suitable. 197 cedures for the assay of tannins in sorghum forage are much less numerous. However, Cummins (4) and Paroda et al. (10) used the vanillin-HCI procedure of Burns (2), and Gourley and Lusk (6) used that of Maxson and Rooney (9) for estimates of forage sorghum tannins. In some of the cited studies precautions were taken to avoid false positive tests for tannins. For example, Price and Butler (11) and Sarkar and Howarth (13) used controls in which vanillin was omitted from the vanillin-HCI reagent, and EI Tuhami et al. (5) used extracts without either vanillin or HCI as controls. In the cited reports of tannins in sorghum forage, however, it appears that such controls were not used. In preliminary attempts to use a vanillin-HCI procedure to detect tannins in sorghum forage, we observed a false positive reaction for some cultivars, that is, leaf extracts from these cultivars produced a red color in the presence of HCI with or without vanillin. This paper presents the results of a series of experiments in which these false positive tests were investigated. MATERIALS AND METHODS Blades of upper leaves of field-grown forage sorghum plants were used in these experiments. Most of the plants were at or just beyond the stage of panicle emergence when sampled. Leaves were dried at about 75 C and were then ground in a Wiley3 mill to pass a I-mm screen. The ground tissue was stored in tightly covered jars in the dark at room temperature until used. The vanillin-HCl assays described by Burns (2) and Maxson and Rooney (9) served as the basis for the procedures used in this study. The Maxson and Rooney procedure differs from that of Burns in that 1 % concentrated HCl in methanol rather than pure methanol is used as the extractant. Unless otherwise indicated, leaf extracts were prepared by placing ground leaf tissue in test tubes or flasks, adding methanol, capping the vessels, and shaking the mixtures occasionally over a period of 16 to 24 hours at room temperature. Filtrates or supernatant solutions from these extractions were used in the assays. Further procedural details are given in connection with specific experiments. Reagent grade HCl and organic solvents were used in all experiments. Vanillin and D-catechin were obtained from the Sigma

Recent emphasis on energy problems has stimulated efforts to identify crops capable of producing high yields of biomass that can be converted into ethanol. The sorghum [ Sorghum bicolor (L.) Moench] crop includes cultivars and hybrids that vary widely in the relative amounts of grain and stalks produced, and also in the sugar content of the stalks. N39 × Wray, a relatively sweet hybrid with good grain potential, and Wray, a cultivar with sweet stalks, were compared for their grain and sugar yields, alcohol production potential, and other related characters. The hybrid produced heavier seeds, more seeds per head, and more heads per ha than the Wray. This resulted in a grain yield of 6630 kg/ha, about 3.4 times that of Wray. Wray produced about 10% more biomass with the stalk sugars accounting for about 80% of the total alcohol potential as compared to 45% for the hybrid. The total alcohol production potential (stalk plus grain) was 1.11 times as great for the hybrid as for Wray.

This study was conducted to evaluate a wider range of plant types and agronomic and quality traits than had been previously reported for blends of sorghum [Sorghum bicolor (L.) Moench]. Three groups of sorghum cultivars and hybrids, each including a i-dwarf (tall), a 2-dwarf (medium), and a 3-dwarf (short) sorghum, were used in a 2-year study. Within each group the following seven blends were compared: tall (T), medium (M), and short (S); T and M; T and S; M and S; T alone; M alone; and S alone. Principal emphasis was given to total dry matter (DM) production (sum of leaves, stems, and heads). It was concluded that blends consisting of entries differing in stature had no significant DM yield advantage over pure stands of T types. Yields of leaf and stem digestible dry matter and crude protein were closely correlated with leaf and stem DM yields. Yields of DM per stalk for S plants were usually highest when these plants were grown in pure stand, but DM yields/stalk for M and T entries were generally highest when these entries were grown in blends. Additional index words: Sorghum bicolor (L.) Moench, Forage yield, Crude protein, Digestibility, Intercropping. EX: ENSIVE .studies on. the theory and practice of many kmds of mtercroppmg have been published (4), but little published information is available on the intraspecific intercropping of sorghum [Sorghum hicolor (L.) Moench] cultivars and hybrids. In a 5-year study at Hays, Kansas, Ross (6) compared the yields of five grain sorghum hybrids with yields of all possible 1: 1 blends derived from them. No consistent advantage in grain yield was observed for any of the blends. Reich and Atkins (5) studied the performance of grain sorghum parental lines and hybrids, and two-component parental blends and hybrid blends in nine Iowa environments over 2 years. Averaged across all environments, most of the blends produced somewhat more grain than the mean of pure stand yields of their components. In each environment except one, however, the highest yield was produced by a hybrid grown in pure stand. More recently Skidmore and Hagen (7) studied the response of 2-dwarf and 3-dwarf isogenic hybrids of 'RS702' grain sorghum grown with the taller 2-dwarf hybrid as a shelter for the 3-dwarf hybrid. They observed that yields of grain and forage per culm of the 2-dwarf hybrid were higher in blends consisting of 13 or 25 % 2-dwarf seed than in a pure stand of the 2-dwarf hybrid. However, for the 3-dwarf hybrid yields of grain and forage per culm with this hybrid comprising 87 or 75% of the blend were not significantly different from yields per culm in a pure stand. The cited studies of Ross and of Reich and Atkins employed only grain sorghums; that of Skidmore and Hagen involved a single 2-dwarf, 3-dwarf pair of entries. In the study reported in this paper a wider range of plant types was utilized, and a more extensive list of traits was examined. MATERIALS AND METHODS Three groups, each consisting of three different entries, were included in this study. One of the entries in each group was a 1dwarf (tall) forage sorghum cultivar or hybrid; one was a 2-dwarf (medium) hybrid resulting from crosses involving germplasm from 'Early Hegari' and the forage sorghum cultivars 'Atlas', 'Rox', 'Sart', and 'White Collier'; and the third was a 3-dwarf grain sorghum hybrid (Table 1). Entries were chosen such that in each group the short (S) entry matured before the medium (M) entry which matured before the tall (T) entry. Within each group seven blends were prepared (Table 2). Blend trns, for example, consisted of a blend of the T, M, and Sentries, tm was a blend of T and M, etc. In some of the analyses the 12 individual components of the blends were considered. The relationship of these components to the blends also is shown in Table 2. The 'Contribution from USDA-ARS, and the Nebraska Agric. Exp. Stn., Lincoln. Published as Paper No. 6143, Journal Series, Nebraska Agric. Exp. Stn. Received 26 Jan. 1981. The work reported was conducted under Nebraska Agric. Exp. Stn. Projects 12-088 and 12-114. 'Supervisory research geneticist, USDA-ARS and professor of Agronomy; and George Holmes professor of Agronomy; Univ. of Nebraska, Lincoln, NE 68583, respectively. 224 CROP SCIENCE, VOL. 22, MARCH-ApRIL 1982 t Two-year means. entries grown in pure stands. Table 1. Groups of entries used in the sorghum blend experiment.

The concentrations of minerals in forages are important in satisfying animal requirements, but little attention has been given to determining these concentrations in forage sorghum [ (L.) Moench.] breeding programs. In this 2-yr study, the objectives were to determine the contents, genetic variability, combining ability effects, and correlations for 12 mineral elements in 49 experimental hybrids of forage sorghum. Mineral elements studied were N, Mg, Si, P, S, Cl, K, Ca, Mn, Fe, Cu, and Zn. General combining ability (GCA) effects exceeded specific combining ability (SCA) effects for all elements except P, Cl, and Fe in females, and S and Cl in males. The GCA and SCA effects in females were low for both Fe and Cu. Genetic ratios, resembling heritability, were higher for GCA than SCA except for Cl and Fe in females, and S and Cl in males. Thirty phenotypic correlations among the 12 elements were statistically significant. Some of the highest values were from the relationships of N with P, S, Cu, and Zn; Mg with Si and Cu; Si with Ca; P with Cu and Zn; S with K; and Cu with Zn. Some of the highest phenotypic correlations calculated among the mineral elements and 12 agronomic and quality traits were the positive association of protein with P, and the negative relationship of height with N and P, Brix of stem juice with K, and neutral detergent fiber with P. Data obtained in these studies show the feasibility of altering the mineral content of forage sorghum by breeding.

The variation among 49 Fl forage sorghum [Sorghum bicolor (L.) Moench.] hybrids from a 7 X 7 cross-classified design was explored in 1979 and 1980 for the following silage traits: dry matter (DM), crude protein, in vitro dry matter disappearance (IVDMD), neutral detergent fiber, acid detergent fiber, acid detergent lignin, ammonia, lactate, and Brix of the juice from fresh stalks. Wider ranges generally were found for male than for female parental means. Means for most traits were significantly different among entries. Significant differences among hybrid means over males and over females were found for only DM, IVDMD, and Brix. Interactions with years existed for most traits. Genetic ratios calculated from the mean squares indicated that general combining ability was important for DM, IVDMD, and Brix. Simple correlation coefficients between traits measured on silage and on fresh-dried samples from the same hybrids were all significant. In view of the effort required to make and evaluate silage samples, initial selection for traits used to estimate quality in fresh-dried samples appears to be the best approach for improving the quality of forage sorghum silage. Additional index words: Sorghum bicolor (L.) Moench., Protein, IVDMD, Fiber, Lignin, Ammonia, Lactate, Brix, Combining ability. T HE inheritance of several constituents used to estimate quality in dried samples from freshly harvested forage sorghum [Sorghum bicolor (L.) Moench.] hybrids was reported in an earlier paper (19). Forage sorghums often are preserved and utilized as silage (18), and some constituents used to estimate quality have been shown to be altered during the ensiling process (5). Information about the inheritance of silage constituents is needed in forage sorghum breeding programs. The large number of silage samples required in inheritance and breeding studies necessitates the use of simple, economical, miniature silos. Sealed glass canning jars meet this requirement and have been used experimentally for more than 50 years (1, 6, 11). It is recognized that the silage produced in miniature silos may differ from that produced in field scale silos, but small silos provide a feasible means of obtaining reasonable estimates of silage quality (16) from a large number of entries. Some factors that can affect silage quality include dry matter percentage at time of ensiling, lactic acid concentration, ammonia content, and availability of fermentable carbohydrates. Many traits such as in vitro dry matter disappearance (IVDMD), crude protein (CP), dry matter (DM) percentage, neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL), can be used to estimate quality in both freshly harvested sorghum forage and silage. Ward et al. (24) found that lower DM percentage of sorghum silage resulted in lower DM intake. However, this relationship was probably secondary since Thomas et al. (22) demonstrated that a change in the DM content of a silage at the time of feeding did not alter the DM intake. The intake depression associated with higher moisture silages may be due to their increased levels of organic acids, particularly lactic acid (9). Thomas et al. (22) reported a reduction in dry matter intake of silage when lactic acid was added to the silage or introduced directly into the rumen. McLeod et al. (15) found that increases in pH of the silage increased DM intake while decreases in pH reduced DM intake. The breakdown of proteins to ammonia is considered detrimental to silage quality (3) because palatability is reduced and some of the ammonia is lost through volatilization as the silage is fed. Some workers also have associated higher soluble N content with lowered intake of silage (17). McCullough and Cummins (14) reported that sorghum silages are deficient in fermentable carbohydrates, the energy source required for the desired lactate-type ensiling process. Similarly, Zimmer (25) stated that low sugar con1 Joint contribution of the USDA-ARS and the Nebraska Agric. Exp. Stn., Lincoln. Published as Paper No. 6783, Journal Series, Nebraska Agric. Exp. Stn. Received 12 Feb. 1982. The work reported was conducted under Nebraska Exp. Stn. Projects 12-088 and 12-114. 2 Formerly graduate research assistant (now assistant professor of agronomy, Auburn Univ., Auburn, AL 36849); George Holmes professor of agronomy; supervisory research geneticist, USDA-ARS; and associate professor of animal science, Univ. of Nebraska, Lincoln, NE 68583 respectively. PEDERSEN ET AL.: ESTIMATING SILAGE QUALITY OF SORGHUM HYBRIDS 377 centrations contributed to a less desirable heterofermentive ensiling process. However, Gourley and Lusk (8) proposed that if more soluble carbohydrates are available than the 6 to 8'70 required for fermentation, the excess may be metabolized to CO2 and water. They also stated that sweet sorghum silages generally are inferior to intermediateand graintype silages in terms of feeding value. The objectives of this study were to determine the extent of variation among 49 experimental forage sorghum hybrids for several constituents used to estimate silage quality and to estimate the proportion of the total variation that was caused by genetic variation. In addition, general combining ability (GCA) and specific combining ability (SCA) effects were estimated. MATERIALS AND METHODS The 49 Fl hybrids used, growing conditions, harvesting and sampling techniques, and statistical and genetic analyses were described in an earlier paper (19). Pertinent information relative to this study is repeated here. The 49 hybrids were produced by crossing each of seven male-sterile lines ('Redlan', 'N35', 'N38', 'N48', 'N4692', 'KS5', 'N5013'3) to each of seven pollinator lines ['Early Hegari-Sart' (EH -SAR T)3, 'Early Hegari-White Sourless' (EH-WS)3, 'Early Hegari-Rox' (EH-Rox)3, 'N6229', 'Rox', 'White Collier' (WC), and 'H60-29'3]. The 49 hybrids were grown in a randomized complete block design with three replications at the University of Nebraska Field Laboratory, Mead, in 1979 and 1980 using standard agricultural practices for that area. Plots were three 9.14-m rows spaced 0.76 m apart with plants hand-thinned to a spacing of approximately 15 cm. A 4.75-m section of the middle row of each plot was harvested after all entries had reached physiological maturity (24~ 27 Sept. 1979 and 6~ 7 Oct. 1980). Random subsamples were passed through a small chopper, mixed thoroughly, and packed as tightly as possible into the miniature silos which consisted of O.95-liter glass jars. The jars were immediately sealed with new canning lids. The lids and jar tops were dipped in melted paraffin at the end of each day to ensure an airtight seal. The miniature silos were then incubated at 28 C to promote a lactate-type fermentation (13). After 4 weeks the ensiling process was assumed to be complete, the jars were opened, and subsamples were withdrawn for drying. The jars were again closed, and jars and contents were held in a freezer until additional subsamples were withdrawn for the preparation of extracts. The subsamples withdrawn for drying were weighed, dried to a constant weight at 57 C in a forced-air oven, and reweighed. Data were collected for the following constituents used to estimate quality: Percent DM. Calculated from the difference in weights of wet and dried silage samples. Percent CP (Percent N X 6.25). Determined by the Kjeldahl procedure (10). IVDMD. Determined by the two-stage technique of Tilley and Terry (23) and expressed as a percentage. Percent NDF. Determined by the high concentrate procedure of Robertson and Van Soest (20). Percent ADF, ADL. Determined by the detergent fractionation procedures of Goering and Van Soest (7). Extracts were prepared from separate subsamples of the wet silage using a modification of Byer's (4) extraction procedure for organic acid analysis in fermented feeds. Twenty-five grams of silage, 70 ml of O.OlN H 2S04, and a thymol crystal to prevent bacterial growth, were placed in a 125-ml Erlenmeyer flask, mixed, and refrigerated for 24 hours to equilibrate. The liquid was filtered 3 Experimental forage sorghum lines. through Whatman #40 filter paper, and the filtrate was frozen and thawed once to aid in precipitation. Four milliliters of the thawed suspension was placed in a centrifuge tube with 1 ml of 10% (w Iv) trichloroacetic acid and centrifuged for 15 min at 12,000 X g. The supernatant was decanted and frozen for later analyses of lactate and ammonia. The determination of lactic acid content involved the use of Barker and Summerson's (2) co:orimetric technique, and ammonia content was determined by an adaptation of the indophenol method of McCullough (12). Measurements of Brix from three stalks/plot were made using a hand-held refractometer on 17 and 18 Sept. 1979 and on 9 Oct. 1980. Juice from the fifth internode, counting the peduncle as the first, was squeezed onto the refractometer stage for these readings. The genetic analyses of these data followed the method described by Ross et al. (21). Although described in an earlier paper (19), the procedure is repeated here for convenience. The entry source of variation was partitioned into females, males, and females X males, and the entry X year interactions were partitioned into females X years, males X years, and females X males X years. Mean squares (MS) were equated to their expected values and solved for components estimating the variance among the fixed effects in the corresponding sources of variation. F -tests were made for females as MSr/MSfy , for males as MSm/MSmy, for females X males as MSfm/MSfmy, and for females X males X years as MSrmy/MSe. Genetic ratios were estimated as follows: (lT~/RFY + lTfmy/FY + 8fm/F + IT;;'y/Y + 8;;')

The influence of autoclaving and of fumigation with eight chemical formulations upon microbial numbers in straw-amended Peorian loess and the extent to which these treatments affect the degree of aggregation brought about by subsequently introduced inocula were determined. A mixed soil flora, Stachybotrys atra and a blank inoculum consisting of sterile salt solution, were compared. Only in the autoclaved soil was a mixed soil flora inoculum consistently superior to the blank inoculum in promoting aggregation of the Peorian loess. It was found that inoculation with S. atra could greatly increase the aggregation of fumigated Peorian loess in treatments where the fumigation was effective in reducing microbial numbers to a low level, and where the fumigant exerted little if any residual toxic effect upon the S. atra inoculum. Most promising of the fumigants tested were chloropicrin and Telone. The aggregation brought about by inoculation with S. atra in Peorian loess fumigated by appropriate amounts of either of these chemicals was approximately 'Contribution by the Soil & Water Conservation Research Division, ARS, USDA, and the Department of Agronomy, Nebraska Agr. Exp. Sta., Lincoln, cooperating. Published ~ith the approval of the Director as Paper No. 851, Journal Senes, Nebraska Agr, Exp. Sta. Presented before Div. III, Soil Science Society of America, Nov. 20, 1957, at Atlanta, Ga. Received Dec. 12, 1957. Approved April 8, 1958. 'Bacteriologist, Soil & Water Conservation Research Division, ARS, USDA; and Associate Professor of Agronomy and fanner Graduate Student, Univ. of Nebraska, respectively. two-thirds as high as that produced by the same organism following autoclaving of the soil material. SOI L STRUCTURE influences water intake, soil aeration, and degree of erosion by wind and water and th~reby affects the growth of microorganisms as well as higher plants. The structural status of a soil is the net result of many complex reactions involving physical, chemic~l, and biological agents. Among the biologIc~1 agents, mIcro?~­ ganisms are of great importance :in soil structure stabilization. Microorganisms, during decomposition of plant and animal residues, produce numerous organic compounds and release inorganic substances, both of which may affect stability of the soil units in various ways (7). Some of the compounds produced and released exert a tremendous effect on stability, while other compounds have only a slight effect or no effect at all. Some of the natural products produced by microorganisms are more effective per unit weight than are the synthet~c sta?!lizers (2, 9). Microorganisms vary greatly in then ability to stabilize the soil (8). Some, such as the cellulose decomposer Stachybotrys atra, are highly effective in increasing soil aggregation (3, 4). . Previous experience has shown that before an organism such as S. atra can be used as an aggregator, ;.t is necessary that the natural microflora be reduced to a low point. This reduction can be accomplished by heat treatment, such as autoclaving. Soil fumigation to combat plant pathogens is becoming an increasingly common agro~­ omic practice (5, 10). The chemicals may kill saprophytic Published in Soil Science of America Proceedings (1958), vol. 22

SynopsisFree and bound coumarin are extracted by autoclaving leaf tissue in water. Free coumarin is determined directly from an aliquot of the water extract diluted in alkali. Additional autoclaving of the extract in concentrated alkali is required for release of the coumarin from the bound form, permitting an assay for total coumarin. Bound coumarin values are then obtained by difference. Results of the assay as applied to four different phenotypes of sweetclover are given.

Seed size has been shown to have an important In­ fluence on the emergence and early growth of seedlings of several forage legumes, but only fragmentary infor­ mation on seed size effects in sweetclover (Melilotus spp.) has been published. Therefore, a 2-year study was con­ ducted in which spring and fall seedings of small, medium, and large seeds of 'Madrid' and 'N13' sweetclover (Meli­ lotus officinalis (L.) Lam.) were made at depths of 19, 38, and 57 rom. The spring seeding also included a com­ parison of a companion crop (oats) with no companion crop. Data on emergence score and plant height were collected for the spring seeding only; stand count and dry matter yield of tops and roots were measured for both spring and fall seedings. The companion crop had little effect on emergence score, but it reduced stand count and plant height, and was highly detrimental to dry matter yield. The perform­ ance of Madrid was poorer in all respects than that of N13, an experimental cultivar with relatively large seeds. As planting depth was increased, stand counts decreased, but the performance of plants that emerged from the greater depths was relatively good. The most striking results of increased seed size appeared to be improved emergence score and increased dry matter production.

Information about the extent of variation in quality traits among plants sampled at the same developmental stage but on different dates would be useful to forage researchers. The primary purpose of this study was to obtain such information for five forage sorghum [Sorghum bicolor (L.) Moench] cultivars. Percent dry matter, crude protein, and in vitro dry matter disappearance of leaf, stem, and whole plant samples were determined for field-grown samples harvested on three different days during early head emergence (EHE) and once during physiological maturity (PM) for each cultivar. Orthogonal comparisons between EHE and PM sampling stages and among the· EHE samples were made. Results of the 2-year study indicated that statistically significant differences existed among EHE samples and between EHE and PM samples. However, differences among EHE samples were generally much smaller than those between EHE and PM samples and may be of little importance for some applications. Additional index words: Protein, Digestibility, Dry matter, IVDMD, Sorghum bicolor. DIFFERENCES in quality traits of forage sorghum [Sorghum bicolor (L.) Moench 1 sampled at various stages of development or under different management regimes are well documented (1, 3, 5, 6, 9). Much less information is available, however, on possible differences among plants of the same genotype and within the same field plot that reach a given developmental stage at different ages in terms cf O:3.ys after planting. It would be useful for forage sorghum researcht, to know whether the quality traits of a plant that reaches a specific developmental stage on a certain day provide an accurate indication of the traits of other plants in the plot as they reach this stage several days later. An easily identified stage in the development of forage sorghum is the end of the boot stage which is signaled by the beginning of head emergence. This study was designed to determine the extent of variation in percentages of dry matter, crude protein, and in vitro dry matter disappearance occurring among forage sorghum plants that reached early head emergence at different times after planting. Plants also were sampled at physiological maturity for comparison with the samples taken at head emergence. Materials and Methods Five forage sorghum cultivars ('Atlas', 'Brawley', 'Coleman', 'Early Hegari-Rox', and 'White Collier') were grown in 1976 and 1977 in a randomized complete block design with three replications at the University of Nebraska Agronomy Farm, Lincoln, Nebr. 1 Contribution from the USDA-ARS, and the Dep. of Agronomy, Nebraska Agric. Exp. Stn., Lincoln. Published as Paper No. 6861, Journal Series, Nebraska Agric. Exp. Stn. Received 21 Apr. 1982. 2 Former graduate research assistant, Univ. of Nebraska, now assistant professor of agronomy, Auburn Univ.; George Holmes professor of agronomy; and supervisory research geneticist, USDA-ARS, and professor of agronomy, Univ. of Nebraska, Lincoln, NE 68583. 3 Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the USDA or the Univ. of Nebraska, and does not imply its approval to the exclusion of other products that may also be suitable. Each plot consisted of five 6.7-m rows spaced 0.76 m apart. Planting dates were 5 May 1976 and 16 May 1977. An application of 112 kg N/ha was disked into the soil [Kennebec silt loam (Cumulic Hapludoll) 1 prior to each planting, and plantings were followed immediately by applications of a mixture of propachlor (2-chloroN-isopropylacetanilide) and atrazine [2-chloro-4-(ethylamino)-6(isopropylamino)-s-triazinel at 3.75 and 1.25 kg a.i./ha. To supplement rainfall, the plots were flood irrigated once, at about midsummer, both in 1976 and 1977. The stage of early head emergence (EHE) was defined as occurring when the very tip of the head was first visible. When at least one culm in each row of a cultivar reached this stage, sampling of that cultivar was initiated. One culm at EHE was sampled from each row of the cultivar on sampling day one (EHE 1). Two and 4 days later, another single culm that had just reached the EHE stage was harvested from each row of the cultivar (EHE 2 and EHE 3, respectively). One set of samples also was taken for each cultivar at physiological maturity (PM) as indicated by black layer formation (4). The five culms harvested from a plot at each sampling day were cut off at the surface of the soil and were separated into leaf blade, stem plus leaf sheath, and head portions. Each culm was treated individually for assay. The immature head was considered to be part of the stem in all EHE samples. The separated samples were weighed and then dried at 75 C in a forced draft oven to constant weight. Dried samples were ground in a Wiley3 mill to pass a 1mm screen, and subsamples were stored for subsequent analysis. Percent crude protein (CP) was determined for each sample as Kjeldahl N X 6.25 (7), and in vitro dry matter disappearance (IVDMD) by a modification of the two-stage technique of Tilley and Terry (8). Percent dry matter (DM) was calculated for each sample, and values for CP, IVDMD, and DM were calculated for the total plant from the values for the leaf blade, stem, and, for PM samples, head portions. Analysis of variance with orthogonal comparisons (2) was completed for each of the traits under study. Results and Discussion Mean squares for some of the sources of variation involved in this study are shown in Table 1. For many of the measured traits, statistically significant variations were associated with year interactions. Therefore, the mean squares are presented on an individual year basis. Inspection of these mean squares shows that significant differences were detected between the EHE and PM stages for all traits except total plant IVDMD in 1977, and that significant (EHE vs. PM) X cultivar interactions were found in all cases except total plant CP in 1976. Significant differences among the three EHE sampling dates were shown for most traits. However, the mean squares for variation among EHE samples were generally much smaller than those for EHE vs PM. Means for each cultivar X sample time X year X trait combination are shown in Table 2. As expected, leaves and stems of all cultivars harvested at EHE were lower in DM than those harvested at PM. The difference between EHE and PM samples was especially pronounced for total plant values, reflecting the relatively high DM percentages of heads at the PM stage (separate data for heads not shown). Decreases in forage CP and IVDMD are generally expected as plants advance in maturity. Comparison of CP values of leaves, stems, and total plants indicates that such a decrease occurred for all cultivars between EHE and PM. Similarly, leaves of all cultivars decreased in IVDMD between these two stages. However, Early Hegari-Rox was

Families of two indiangrass [Sorghastrum nutans (L.) Nash] populations were used in this study with a family plot consisting of a parental clone and four half-sib progeny_ The experimental design was a randomized complete block with two replications. The regression of offspring on the parent in the same plot was used to obtain heritability estimates in which environmental covariances might be expected to inflate the parent-offspring covariance. The regression of offspring in one replication with its parent in another replication, and vice versa, was used to obtain heritability estimates whereby environmental covariances should be minimized. Heritability estimates for heading date and plant height, which had high heritabilities, were similar for the two estimation techniques. Heritability estimates for forage yield and in vitro dry matter digestibility (IVDMD) appeared to be influenced by environmental covariances. Mean heritability estimates for forage yield and IVDMD for the three sets of data analyzed were 0.64 and 0.94 respectively, when parents and offspring in the same plot were used in the regression analyses and 0.28 and 0.42 respectively, when parents and offspring in different plots were used. Heritability estimates for forage protein content averaged 0.49 and were similar for both methods of estimation. Additional index words: Sorghastrum nutans, Forage yield, Forage quality, In vitro digestibility, Genetic covariances. 1 Contribution of the Dep. of Agronomy, Univ. of Nebraska, and AR, SEA, USDA. Published as Paper No. 5821, Journal Series, Nebraska Agric. Exp. Stn. Received 26 Sept. 1979. • Research agronomist, AR, SEA, USDA; George Holmes professor of Agronomy, Univ. of Nebraska; and supervisory research geneticist, AR, SEA-USDA, Univ. of Nebraska, Lincoln, NE 68583. R EGRESSION of offspring on parents or on one parent is a method of estimating heritability commonly used by plant breeders. The covariance between a parent and its offspring produced by random mating is an estimate of one-half the additive genetic variance in a population (3, 5). The mean of several offspring is usually used as the offspring value. The covariance between offspring and parents is divided by the phenotypic variance of the parents to obtain the regression coefficient. Because the covariance between offspring and parent is only one-half the additive genetic variance, the regression coefficient is multiplied by two to obtain an estimate of narrow sense heritability (5). Regression of offspring on parents to estimate heritability is based on the following assumptions (3, 4): a) normal diploid and solely Mendelian inheritance, b) no environmental correlations among relatives, c) population in linkage equilibrium or no linkage among genes controlling the traits studied, d) the relatives are noninbred, e) the genetic population is mating at random. Heritability estimates obtained by parent-progeny regression are often used because the estimates obtained are valid both when the parents are selected on some basis and when they are chosen at random from a population (5). The purpose of this paper is to demonstrate the inflationary effect that environmental covariances might have on heritability estimates if the second assumption, i.e., no environmental correlations among relatives, is violated. In 1973, we began a quantitative genetic study on indiangrass, Sorghastrum nutans (L.) VOGEL ET AL.: PARENT-PROGENY REGRESSION IN INDIANGRASS 581 N ash. Our results from this study were used to estimate the effect of environmental covariances on heritabilities based on parent-progeny regression. Indiangrass is a warm-season grass native to the prairies and plains of the United States (9) where it is used in pastures and rangelands for summer grazing. Limited genetic research on this grass has been reported although several varieties have been released. In our study we used two varieties that are adapted to the central Great Plains. 'Holt' was developed by mass selection from ecotypes collected in northeast Nebraska while 'Oto' was developed by mass and progeny selection from ecotypes collected in southeast Nebraska and northeast Kansas (7). MATERIALS AND METHODS In 1973. space-transplanted populations of a Holt X Oto synthetic and Oto indiangrass were sampled at panicle emergence for in vitro dry matter digestibility percentage (IVDMD) and for crude protein. Each nursery contained approximately 1.000 plants_ The Holt X Oto synthetic was established from seed harvested from Holt plants exposed to Oto pollen in an open-pollinated Holt X Oto crossing block. Because Holt flowers 20 days earlier than Oto. it is doubtful if any crossing occurred and the Holt X Oto synthetic will be referred to as Holt. Plants in both nurseries were 2 years old when sampled. The Holt population was located at Lincoln. Nebraska. while the Oto population was located at Mead. Nebr.. which is approximately 55 km northeast of Lincoln. In 1973. seed from open-pollinated plants was harvested from plants sampled from the two populations. In 1974. a twice-replicated. space-transplanted half-sib progeny nursery was established at the Mead Field Laboratory with seedlings grown from open-pollinated seed from 41 Holt and 150 Oto parents that represented the high and low ends of the range in IVDMD. protein percentage. and heading date for the two populations. In 1975. two ramets (propagules) of the parental clones similar in size to the seedlings established the p'revious year were transplanted into the same nursery. A famIly plot consisted of a five-plant row with a parental ramet and four progeny seedlings. The parental ramet was at the end of the plot. Rows and plants within rows were spaced 1.1 m apart. The experimental design was a randomized complete block. Holt and Oto families were randomized together in the ./ same nursery. but because of maturity differences were analyzed .

Youngest fully expanded leaves, from plants of M. officinali$ (L.) Lam. seeded at weekly intervals but harvested on the same day, were approximately 1.5 to 2.0 times as high in content of glucose, fructose, and sucrose at all stages of growth as leaves from comparable plants of M. infesta Guss. These plants varied in age from 3 to 14 weeks. Levels of glucose and fructose increased with increasing plant age in both species. Sucrose content increased in M. officinalis but decreased in M. infesta as the plants matured. When plants of approximately the same age were sampled at six different dates, the level of each sugar again was higher in young leaves of M. officinalis than in corresponding leaves of M. infesta at each sampling date. Evidence reported in this and preceding papers suggests that differences in sugar content may influence the degree of resistance or susceptibility of M elilotus leaves to feeding by the adult sweetc10ver weevil (Sitona cylindricollis Fahraeus), but that these differences apparently are not primarily responsible for the observed differences in resistance displayed by M. officinalis and M. infesta. Additional index words: Sitona cylindricoUis, Sugars, Insect resistance. F by the adult sweetclover weevil, Sitona cylindricollis (Fahraeus) was stimulated by a water-soluble fraction extracted from Melilotus leaves (4, 6). The feeding stimulant, designated Stimulant A, was found in chromatographically purified leaf extracts from the weevil-resistant Melilotus infesta Cuss., as well as from a susceptible species, M. officinalis (L.) Lam. Stimulant A was subsequently fractionated into three feeding stimulants which were identified as glucose, fructose, and sucrose (3). The isolated compounds and corresponding reagent grade sugars were identical in chromatographic and chemical behavior, and also in feeding stimulant activity as measured with the sweetclover root disk bioassay (7). Sucrose stimulated the greatest amount of feeding when the three sugars were compared at equal concentrations in the same bioassay (3). Disks treated with glucose and fructose were fed upon equally at the three concentrations tested. Sweetclover weevil resistance apparently depends upon a balance between feeding stimulants and feeding deterrents; and the relative contributions of the different factors vary considerably with stage of leaf and plant development (1). Information concerning 1 Contribution from the Nebraska Agricultural Experiment Station, Lincoln, Neb., and the Crops Research Division, ARS, USDA. Supported in part by Crops Research Division and Entomology Research Division, Agric. Res. Serv., USDA Grant No. 12-14-100-8027 (33). Published with the approval of the Director as Paper No. 2687, Journal Series, Nebraska Agr. Exp. Sta. Received Dec. 4, 1969. 2 Formerly Assistant Professor of Agronomy, University of Nebraska (present address, Great Western Sugar Co., Longmont, Colo.); Research Geneticist, Crops Research Division, ARS, USDA; and Bert Rodgers Professor of Agronomy, University of Nebraska, respectively, Lincoln, Neb., 68503. The technical assistance of Patricia Underwood and Henry J. Stevens is gratefully acknowledged. 477 the relative amounts of the three sugars acting as feeding stimulants in sweetclover leaves would aid in determining the role of these sugars in the mechanisms of resistance and susceptibility of Melilotus plants to weevil feeding. Levels of glucose, fructose, and sucrose in resistant and susceptible plants of various ages are reported in this paper. MATERIALS AND METHODS Plantings consisting of 10 plants of M. infesta (Nebraska strain M70) and 10 of M. officinalis 'Goldtop' (F.C. 38,923) were made in a greenhouse at weekly intervals for 13 weeks, to permit comparison of the sugar content of similar leaves of the two species from plants of varying ages. Natural light was supplemented with cool white fluorescent lamps to provide a 16-hr photoperiod. Sampling and extraction of leaves from plants 3 to 14 weeks of age was accomplished in one afternoon, following !i consecutive sunny days. The youngest fully expanded leaf from each of the 10 plants of similar age and specieS was harvested, and these 10 leaves were bulked. One week after the initial harvest, the newly formed youngest fully expanded leaves were similarly harvested and extracted, again after at least five consecutive days of sunshine. For example, the 10 plants of M. infesta from which the first 8-week sample was taken, were used one week later as the source of material for the second 9-week sample. Thus, there were two samples representing each plant age. Following the harvest of leaves from the 10 plants of a group, the three leaflets of each leaf were removed from the p.etiole. Mid-leaflets, used for the determination of dry matter percentage, were weighed before and after drying at 110 C for 6 hours. The 20 side leaflets were weighed, washed with water, dropped into 10 ml boiling water and autoclaved at 120 C for 20 min. Autoclaved extracts were cooled, leaflets were removed and discarded, and the extracts were stored in a freezer for later assay. Additional plantings of both species were made every 3 to 4 weeks, so that several extractions over a period of 3 months could be made on plants of approximately the same age. Ten plants of each species, from 5 to 8 weeks of age, were sampled at each of six harvest dates. All sampling and extraction was done during the afternoons of days having full sunshine. The youngest fully expanded leaf from each plant was harvested individually. Mid-leaflets were used for the determination of dry matter percentage, while the two side leaflets were weighed, dropped into 2 ml boiling water, and autoclaved at 120 C for 20 min. Extracts were stored in a freezer for later assay. In order to remove compounds interfering with the sugar determinations, crude extracts were partially purified by preparative paper chromatography in a manner similar to that described previously (4, 6). Chromatograms were developed with a solvent composed of isopropyl alcohol, ethyl acetate, and water (8:1:3, v/v/v). Glucose, fructose, and sucrose were confined to a single band extending from Rf 0.30 to 0.52; this band was eluted with water. Thin-layer chromatography of the eluates, using the procedure described by Akeson et al. (3), indicated that glucose, fructose, and sucrose were the only sugars present, and that other interfering reducing compounds had been effectively removed. Eluates were assayed for glucose and fructose content by means of a modification of the colorimetric method described by Ting (9). Total reducing sugars were determined by first mixing 1 ml of eluate (representing 1 to 3 mg of dry leaf tissue) with I ml of alkaline ferricyanide solution in an 18 X 150 mm test tube and heating the mixture in a boiling water bath for 10 min. The tube was then cooled in cold water, and 2 ml of 2 N sulfuric acid and 0.8 ml of arsenomolybdate solution were added. Absorbance of the solution was read at 515 nm with a blank consisting of a similarly treated eluate from a chromatogram to which no extract had been applied. Apparent fructose content was determined by the same procedure described for CROP SCIFNCE, VOL 10, SEPTEMBER-OCTOBER 1970 'olal reducing sugars, except that the mixtures of cluate and ferricyanide were incubated at tiC, C [or 30 min, Clu('ose and frucu;se contents were calculated from the total reducing sugars and apparent fructose values by the equation of Ting (~l), A modification of the resorcinol procedure described by Kulka (8) was used to determine sucrose. One ml of cluate was mixed with 3 ml of 30% HCI (containing 0.216 g/Iiter l"eNH4 (S04)" 12 H 20) and 3 ml of O'()5% resorcinol (w/v in absolute ethanol), The mixture was heated at 73 C [or I hour, cooled, and read at 480 nm, The reading obtained provided a measure of total fructose residues (free fructose plus the fructose component of sucrose), Calculation of sucrose content was based on the difference between total fructose residues and the actual fructose content (obtained from the reducing sugar procedure described in the preceding paragraph), , The efficiency of the above procedures was studied III preliminary experiments involving' recovery of glucose, Iructose, and sucrose added to crude extract. Recoveries were 95 to 99% lor glucose, 93 to 94% for fructose, and 98 to 100% for sucrose, RESULTS AND DISCUSSION Average contents of glucose, fructose, sucrose, and total sugars (sum of glucose, fructose, and sucrose) in the youngest fully expanded leaves of iVL mfcsta and J\!I, otticinaZis plants [rom 3 to 14 weeks of age are presented in Fig, 1, Each value in the graph represents the mean of two extracts (one extract from each of the two samples representing each plant age), Triplicate chromatograms of each extract were prepared, developed, and assayed, At all ages, the content of each of the sugars was consistently higher in leaves of J\!I, offirinalis than in corresponding J\!I, infcsta leaves, In AI, officinalis leaves, glucose, fructose, sucrose, and total sugar contents, averaged for all plant ages, were 1.8, lA, 1.9, and 1.7 times as high, respectively, as the averages [or corresponding lV[, infcsta leaves, In addition, the content of each sugar, with the exception of sucrose in J\!I, infesta, tended to increase with increasing plant age, The contents of glucose and fructose were higher in leaves of older lV[, infesta plants than in leaves of young plants of lV[, ottirinalis, The sucrose content of lV[, infest a increased slightly as the young plants grew, remained about the same during the intennediate period of growth, and then decreased rather sharply as the plants matured. The contents of sugars from plants of approximately the same age harvested at six different dates are

SynopsisPeroxidase activity in extracts of underground buds of ironweed and leafy spurge was at the lowest level during the period of dormancy in both species, and increased when dormancy was broken. Polyphenoloxidase activity of bud extracts was high for ironweed and very low for leafy spurge, and showed no relationship with dormancy in either species. Ninhydrin-positive substances generally and sulfur-containing amino acids in particular increased during the dormant period.

279 Red clover (Trifolium pratense L.) varieties in the third season of growth, that had been exposed to a heavy infestation of larvae of the clover leaf weevil [Hypera punctata (Fabricius)], differed significantly in percentage defoliation, feeding injury score, plant height, and forage production. Based on these four characters, 'Mammoth,' 'Lakeland,' 'Dollard,' and 'LaSalle' were highly resistant to larval feeding; 'Kenland,' 'Common,' and 'Pennscott' were highly susceptible. Long term second·season yield tests including six of these varieties indicated no con· sistent superiority of any of the varieties in the absence of weevil feeding. -------------------Additional index words: Trifolium pratense, Hypera punctata, Insect preference. -------------------T HE clover leaf weevil [Hypera punctata (Fabricius)] can injure clovers (Trifolium spp.) and alfalfa (Medicago sativa L.), but rarely causes the loss of an entire crop. This insect was first reported in New York in 1880 (4) and has now spread over the United States and Canada wherever clover and alfalfa are grown. Damage to the crop is caused by larval feeding and is most apparent in spring. Smooth-edged notches are eaten out of the sides of the leaves or occasionally, whole leaves are consumed. The larvae feed mainly at night and remain well-hidden during the day in trash on the surface of the ground or at the base of the plants. Generally, larval populations are greatly reduced by a fungus [Entomophthora (=Empusa) sphaerosperma Fresenius]. Infected larvae are yellow to gray instead of the normal green, and can be found curled on the edge of leaves because, unlike normal larvae, they do not drop to the ground during the day (2). Breeding for insect-resistant varieties is particularly appropriate in forages because of their comparatively low per hectare value, the relatively high cost of chemical applications, and the possibility of insecticide residues. Thus, selection of red clover (Trifolium pratense L.) varieties, that are resistant to the clover leaf weevil, would increase the dependability and yield of this important hay and pasture crop. A recent review of insect resistance in clovers (3) revealed no published accounts of resistance to the clover leaf weevil. One reason for the lack of such accounts may be the rarity of serious infestations of this insect. The present paper reports the results of a study on the response of 10 red clover varieties and strains to a heavy natural infestation of the clover leaf weevil that occurred in 1971. MATERIALS AND METHODS Ten varieties and strains of red clover were seeded in 1.5 X 7-m plots with a companion crop of oats (Avena sativa L.) at the Nebr. Agr. Exp. Stn. Field Lab. at Mead, Nebr. on Apr. 24, l Contribution from the ARS, USDA, and the Nebr. Agr. Exp. StlI., Lincoln. Published as paper no. 3791, journal series, under project 12-27, Nebraska Agr. Exp. Stn. Received Oct. 4, 1974. 2 Research geneticist, ARS, USDA; research entomologist, ARS, USDA; and Bert Rodgers Professor of Agronomy, U. of Nebr., Lincoln, NE 68503, respectively. 280 CROP SCIENCE, VOL. IS, MARCH-APRIL 1975 1969. Seven rows were hand-seeded in each plot. The test was arranged in a randomized, complete-block design with four replications. Stands of all entries were excellent. The companion crop of oats was combined on July 15, but the red clover was not harvested during the first season of growth. Second-season growth, harvested from all plots on June 8 and July 15, 1970, indicated no significant differences in yield among the 10 entries. In late May and early June 1971, a heavy natural infestation of clover leaf weevil larvae was noted in this nursery. Striking differences in reaction to feeding by the larvae were noted among the various varieties and strains in the test. Stands of all entries in this third-year nursery remained excellent, and no injury from disease was apparent at the time of this insect infestation. These conditions presented a rare opportunity for a preliminary assessment of resistance to this insect. Consequently, on June II each plot was visually scored for percentage defoliation, and an injury score also was assigned. The latter score was based on a scale of I to 5 (I = no feeding damage observed; 5 = heavy feeding damage). Thus, two scores were assigned for essentially the same characteristic, the relative extent of feeding damage by the clover leaf weevil larvae. Height also was measured on June II immediately before a 0.9 X 6.l-m portion was harvested from the center of each plot to estimate yield. At the time of this harvest, all entries except 'Mammoth' were at 50% bloom, the normal stage for harvesting red clover yield tests. Mammoth had not yet started to flower. A sample of the harvested forage from each plot was used to determine moisture percentage. In addition, after they were dried and weighed, the samples of each entry from the four replications were composited and analyzed for N content by the Kjeldahl procedure. Stand percentage was determined for each plot by approximate measurements of missing portions of each harvested row. RESULTS AND DISCUSSION Performance of varieties and strains of red clover that were subjected to heavy feeding by clover leaf weevil larvae is shown in Table 1. Entries did not differ significantly in percentage stand; as already noted, all stands were excellent. Significant differences among entries were observed for percentage defoliation, injury score, plant height, and forage production. No significant differences were observed in percentage dry weight. Nitrogen percentages (one determination/ entry) varied from 2.33 to 2.6J. On the basis of defoliation percentage, the entries most resistant to the clover leaf weevil were Mammoth, 'Lakeland,' 'Dollard,' and 'LaSalle,' 'Kenland,' 'Common,' and 'Pennscott' were the most susceptible. Classifications of the entries according to injury score, plant height, or yield gave generally similar rankings. The following highly significant correlation coefficients were calculated: percentage defoliation and forage yield, -0.915; percentage defoliation and injury score, 0.975; percentage defoliation and height, -0.829; injury score and forage yield, -0.946; injury score and height, -0.855; and forage yield and height, 0.912. Unfortunately, no third-season red clover stands were available in a noninfested nursery in 1971; thus, a study of performance of directly comparable plant material in the presence and absence of weevil infestation was not possible. However as already indicated, second-season yields from this same nursery in 1970 revealed no significant differences among entries. Furthermore, it is noteworthy that in annual replicated trials at Lincoln or Mead during the years 1959 through 1970 (a period when no clover leaf weevil Table 1. Performance of red clover varieties following infesta· tion of third·season growth by clover leaf weevil larvae. Each figure is the mean of four replications.

The purposes of this study were to determine if phenotypic selection for in vitro dry matter digestibility (IVDMD) could be used to improve the IVDMD for switchgrass tPanicum virgatum L.) and to obtain heritability estimates of IVDMD in switchgrass. In 1973 a selection nursery of over 2,200 plants was established with transplanted seedlings of two similar Nebraska experimental strains. In 1974 about 800 plants that appeared to be agronomically superior were sampled for IVDMD. Single-plant selections for high and low IVDMD were made in each of the nursery rows. In 1976, ramets from 25 of the high-IVDMD selections and from 25 of the low-IVDMD selections were transplanted to establish isolated highand lowIVDMD polycross (PC) nurseries. Seed harvested from these nurseries in 1977 and from the same openpollinated (OP) clones in the selection nursery in 1976 was used in 1978 to establish a replicated seeded sward nursery in which the following strains were compared: high IVDMD PC, high IVDMD OP, 'Pathfinder,' low IVDMD OP, and low IVDMD PC. Forage yields and IVDMD were measured in 1978 and 1979. Over the 2 years the strains averaged 51.5, 50.0, 49.4, 48.3, and 48.1% IVDMD, respectively. The high IVDMD PC strain was significantly higher in IVDMD than the other strains in both years. Realized heritabilities for high and low IVDMD were 0.59 and 0.55, respectively. Strains did not differ significantly for forage yield in either year; thus, the gain in IVDMD by breeding was achieved without any yield loss. Additional index words: Panicum virgatum (L.), IVDMD, Phenotypic selection. SW ITCH G R ASS , Panicum virgatum L., is a warmseason, cross-pollinated perennial grass that is native to the prairies and plains of North America where it is used in pastures and rangelands for grazing and prairie hay. Eberhart and Newell (1959) and Newell and Eberhart (1961) reported that substantial genetic variability existed between and within endemic strains of switchgrass for seed quality, leaf rust reaction, leaf height, plant height, seed yield, plant yield and forage quality. Heritability estimates for visually rated forage quality ranged from 0.27 to 0.70 (Newell and Eberhart, 1961). I Contribution of the Dep. of Agronomy, Univ. of Nebraska, and AR, SEA, USDA. Published as Paper No. 5943. Journal Series, Nebraska Agric. Exp. Stn. Received 16 May 1980. 2 Research agronomist, AR-SEA/USDA; George Holmes Professor of agronomy, Univ. of Nebraska; and supervisory research geneticist. ARSEA/USDA, Lincoln, NE 68583. 3 Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the USDA and does not imply its approval to the exclusion of other products or vendors that may also be

Switchgrass (Panicum virgatum) is a native, perennial warm-season grass commonly used as summer pasture. Three strains of switchgrass with different in vitro DM disappearance (IVDMD) values were evaluated with yearling cattle (Bos taurus) in three replicated grazing trials All strains were similar in genetic background, maturity and yield potential. 'Trailblazer' (high-IVDMD) resulted in higher (P < .05) ADG than 'Pathfinder' and 'low-IVDMD'. Available forage of Trailblazer had higher IVDMD than Pathfinder and low-IVDMD. The IVDMD differences among strains were similar throughout the season; IVDMD declined between .3 and .5 percentage units per day. Forage availability, CP, NDF and ADF, permanganate lignin and ash were similar among strains. This study documents that plant breeders can select switchgrass plants for higher IVDMD and that strains consisting of progeny of these plants can result in higher animal gains than strains cons.isting of progeny of unselected or lowqVDMD plants with similar genetic background, forage yield, maturity, morphology and chemical composition. (

Space-planted populations derived from ‘Holt’ and ‘Oto’indiangrass, (L.) Nash, were used to obtain heritability estimates for forage yield, in vitro dry matter digestibility (IYDMD), crude protein, and heading date and to determine their genotypic correlations. Heritability estimates were determined by variance component analyses and by parent-progeny regression. Average narrow sense heritability estimates for the two populations were 0.43, 0.42, and 0.50, for forage yield, IVDMD, and protein, respectively. Narrow sense heritability estimates for heading date were 0.90 or larger. Genotypic correlations of yield and IVDMD were negative for the Holt population but were positive, although low, for the Oto population in 1976. Protein and IVDMD had positive genotypic correlations for both populations. Heading date was positively correlated with yield but was negatively correlated with IVDMD and protein.

Hydrocyanic acid or prussic acid poisoning of livestock by sorghums [Sorghum bicolor (L.)Moenchl and sudangrasses [Sorghum sudanese (Piper) StapfJ is caused by the digestive liberation of hydrocyanic acid (HCN) from the cyanogenic compound, dhurrin [(S)-p-hydroxymandelonitrile ,8-D-glucopyranosideI found in tissue of these plants. Recent research documented that dhurrin is also present in indiangrass [Sorghastrum nutans (L.) Nash] seedlings. The purpose of this study was to determine the hydrocyanic acid potential (HCN-p) of forage from established stands of indiangrass. Five cultivars representative of indiangrass germplasm of the Great Plains were sampled during the growing season for 2 years from 2 sites in eastern Nebraska. The HCN-p of the indiangrass sampled in this study exceeded 750 mg-1 kg dry wt. (dangerous level) only in spring when new growth was 20 cm tall or less. Levels were less than 500 mg-1 kg (safe) when new growth was at least 40 cm tall and were very low (