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Soybean yield is limited primarily by abiotic constraints. No transgenic soybean with improved abiotic stress tolerance is commercially available. We transformed soybean plants with genetic constructs able to express the sunflower transcription factor HaHB4, which confers drought tolerance to Arabidopsis and wheat. One line (b10H) carrying the sunflower promoter was chosen among three independent lines because it exhibited the best performance in seed yield, and was evaluated in the greenhouse and in 27 field trials in different environments in Argentina. In greenhouse experiments, transgenic plants showed increased seed yield under stress conditions together with greater epicotyl diameter, larger xylem area, and increased water use efficiency compared with controls. They also exhibited enhanced seed yield in warm and dry field conditions. This response was accompanied by an increase in seed number that was not compensated by a decrease in individual seed weight. Transcriptome analysis of plants from a field trial with maximum difference in seed yield between genotypes indicated the induction of genes encoding redox and heat shock proteins in b10H. Collectively, our results indicate that soybeans transformed with HaHB4 are expected to have a reduced seed yield penalty when cultivated in warm and dry conditions, which constitute the best target environments for this technology., HaHB4 transgenic soybean plants significantly outyielded controls in warm and dry environments, showing increased seed number, xylem area, and water use efficiency, and induction of genes encoding redox and heat shock proteins.

Summary •Amaranthus tuberculatus represents one of the most dramatic cases of weed invasion documented in the midwestern USA. The species is infamous for evolving resistance to multiple herbicides, and predicting whether these resistances may be transferred to widespread weeds of the Amaranthus hybridus aggregate is a matter of epidemiological concern. Here, we explore the patterns of genetic exchange between Amaranthus tuberculatus and A. hybridus in an effort to understand whether allele introgression occurs throughout the genome and if fecundity penalties are associated with genetic exchange. •We evaluated 192 homoploid BC1s at 197 amplified fragment length polymorphism (AFLP) loci, as well as two loci associated with herbicide resistance: ALS and PPO. We also assessed the fecundity of each genotype by evaluation of seed production or pollen development. •It was discovered that genetic exchange between the species is unidirectional. Whereas A. hybridus alleles transfer with little or no penalty to A. tuberculatus, the reciprocal exchange is significantly distorted and potentially of limited evolutionary consequence. •Our previous hypothesis suggesting unidirectional introgression at ALS owing to circumstantial linkage is now modified to account for the more generalized distortion of genetic exchange observed in this study.

Large proportions of nonhybrid progeny result from controlled crosses between Palmer amaranth and common waterhemp, both dioecious weeds. Agamospermy was proposed as an explanation for this phenomenon, and here we provide evidence in support of this hypothesis. We evaluated 60 nonhybrid offspring from two interspecific crosses, and all plants were females and had DNA content values similar to the female (Palmer amaranth) parent. Among nine hybrids resulting from these crosses, eight were nonviable (lethal or neuter), and only one hybrid allowed for continued gene movement. Cytogenetic evaluation of this hybrid revealed triploidy, further supporting the occurrence of unreduced gametes in these species. In light of this new evidence, we examine earlier data regarding Palmer amaranth by common waterhemp hybridization and suggest some prior conclusions may be premature. Nomenclature: Palmer amaranth, Amaranthus palmeri S. Wats. AMAPAcommon waterhemp, Amaranthus tuberculatus (Moq.) Sauer var. rudis (S...

In the state of Illinois, waterhemp and smooth pigweed are among the worst agricultural weeds. Previous research shows high potential for hybridization between these two species. However, the actual occurrence of hybrids in natural settings is still uncertain. Morphological similarity between hybrids and waterhemp makes field surveys of hybrids difficult to conduct. The main purpose of this study was to characterize the morphology of waterhemp × smooth pigweed F1hybrids, emphasizing evaluation of characters that may allow for hybrid discrimination in fieldAmaranthuscommunities. Concurrently, the study characterized hybrid reproductive fitness, chromosome number, and DNA content. To accomplish this, hybrids were obtained from field crosses. A species-specific polymorphism in theALSgene was used to verify hybrid identity. Significant differences (α = 0.05) between hybrids and individuals of the parental species were observed for five staminate and five carpellate characters. Of these, five characters differentiated hybrids from waterhemp. However, clustering analyses using these characters indicated that morphological differences were not reliable enough, by themselves, for unambiguous hybrid identification. Also, hybrid homoploidy (2n= 32) with respect to parental species excluded chromosome counts in hybridity determinations. However, DNA content analysis may be used for such purpose. Hybrids had an average of 1.21 pg of DNA per 2C nucleus, a value intermediate to that of parental species. Hybrids produced 3.3 or 0.7% the seed output of parental and sibling waterhemp individuals, respectively. Percent micropollen in hybrids was 95-times greater than in parental species. Hybrid sterility appears to be the most reliable feature for hybrid discrimination when conducting field surveys. However, molecular and cytogenetic analyses as employed in this study may be desired for ultimate identity corroboration.

Summary Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of branched-chain amino acids in plants and is the target of several herbicides. ALS inhibitors have enjoyed popularity as herbicides due to numerous attributes, although their current adequacy in weed control programs is hampered by herbicide resistance. Most cases of ALS-inhibitor resistance have resulted from selection of an altered target site. The study herein reports on an alanine by threonine amino acid substitution at position 122 of ALS as the basis for imidazolinone-specific resistance in an A. hybridus population from Illinois. In vitro inhibition of enzymatic activity ( I 50 ) required 1000-fold greater concentration of imazethapyr in the resistant population compared with a susceptible control. This mutation represents the second ALS alteration associated with herbicide resistance in a natural A. hybridus population.

Field studies have established high potential for hybridization between two important and often coexisting weedy species, Amaranthus hybridus and Amaranthus tuberculatus . Prezygotic reproductive barriers between these species are believed to be limited to pollen competition and availability. A greenhouse study showed that a herbicide-resistance gene ( ALS ) from A. hybridus could be introgressed into an advanced A. tuberculatus background (BC 2 ). However, evidence is lacking in support of such transfer in nature. Postzygotic reproductive barriers may minimize, if not preclude, natural introgression. Indeed, A. hybridus × A. tuberculatus hybrids are characterized by reduced fertility and even floral neuterism. The purpose of this study was to assess hybrid fertility in the BC 1 generation and its relationship with genome structure and segregation at ALS. Fertility was assessed by measuring seed output and by pollen evaluation, and segregation at ALS was determined via a molecular marker system. The two parental species have the same ploidy (2n = 32) but differ in DNA content (2C) values, with A. tuberculatus chromosomes being on average 29% greater than those of A. hybridus . Given that most (98%) BC 1 s were homoploid, 2C values were used as indicators of relative genomic constitution. Fertility in the BC 1 generation was greater than that of F 1 s, and 3% of BC 1 s had seed output similar to that of the parental species. Fertility in the BC 1 did not correlate (in a strict way) with reconstitution of parental genomes. Hybrid sterility appeared to be controlled by relatively few loci. Heterozygosity at ALS was negatively correlated with fertility. Also, the A. tuberculatus ALS allele was not observed in the A. hybridus sexual condition, monoecism. Linkage of ALS to a locus associated (directly or via epistasis) with hybrid sterility may explain the fertility penalty observed with ALS introgression. Moreover, this linkage might explain why sequenced herbicide-resistance ALS alleles from sympatric A. tuberculatus and A. hybridus populations show independent evolution.

Several populations of different Amaranthus species have been reported resistant to single or multiple herbicides. Interspecific hybridization among amaranths is hypothesized to contribute to the evolution of herbicide resistance. Although other studies have shown the occurrence of interspecific Amaranthus hybrids, little has been done to establish the likelihood of hybridization under field conditions. The main objective of this study was to determine potential field frequencies of hybridization between tall waterhemp females and smooth pigweed. Field hybridization plots were established during each of two growing seasons. Individuals of the two species were transplanted to field plots and arranged at varying distances from each other. Hybrid progeny were detected using the acetolactate synthase (ALS) gene as a marker. Smooth pigweed parents were homozygous for a herbicide-resistance ALS allele, whereas maternal tall waterhemps were homozygous for a herbicide-sensitive ALS form. Heterozygous interspecific progeny were thus detected by means of herbicide selection. Molecular and cytogenetic tools were used to verify the validity of the data obtained. Averaged among female waterhemp plants and across the two field seasons, hybridization occurred at a frequency of 33%. A single tall waterhemp plant was capable of producing more than 200,000 hybrids, suggesting little if any gametic incompatibility. All flowering hybrids obtained from tall waterhemp females were of dioecious condition and female sex. Observed sexual segregation was consistent with previously postulated chromosomal XY-type system in tall waterhemp sex determination, where males are the heterogametic sex.

Recent studies have confirmed that weedy Amaranthus species are capable of interspecific hybridization, and such hybridization may foster the evolution of herbicide resistance. However, the extent to which hybridization among these species occurs in nature is unknown. The purpose of this study was to determine the frequency under field conditions at which A. hybridus, a monoecious and predominantly self-pollinated species, would be pollinated by A. tuberculatus, a dioecious species. To do this, parents carrying different alleles at the ALS locus, which encodes a herbicide target site, were used. Male A. tuberculatus parents were homozygous for a dominant herbicide-insensitive allele, while A. hybridus parents were homozygous for a sensitive form. Hybrid progeny therefore could be detected via herbicide selection. Mean hybridization frequencies between 0.4 and 2.3% were obtained, depending on the proximity between parents (P=0.02). The robustness of the hybrid selection assay was verified using a molecular marker and DNA content analyses. Using these techniques, more than 99% of the progeny that survived the herbicide were confirmed to be hybrids. Frequencies obtained in this study were many times higher than the generally expected rate of mutation. Therefore, even minimal fertility in hybrid progeny would support the view that hybridization could play a role in adaptive evolution of weedy Amaranthus species.