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Stripe rust is a devastating fungal disease of wheat caused by Puccinia striiformis f. sp tritici (Pst). The WHEAT KINASE START1 (WKS1) resistance gene has an unusual combination of serine/threonine kinase and START lipid binding domains and confers partial resistance to Pst. Here, we show that wheat (Triticum aestivum) plants transformed with the complete WKS1 (variant WKS1.1) are resistant to Pst, whereas those transformed with an alternative splice variant with a truncated START domain (WKS1.2) are susceptible. WKS1.1 and WKS1.2 preferentially bind to the same lipids (phosphatidic acid and phosphatidylinositol phosphates) but differ in their protein-protein interactions. WKS1.1 is targeted to the chloroplast where it phosphorylates the thylakoid-associated ascorbate peroxidase (tAPX) and reduces its ability to detoxify peroxides. Increased expression of WKS1.1 in transgenic wheat accelerates leaf senescence in the absence of Pst. Based on these results, we propose that the phosphorylation of tAPX by WKS1.1 reduces the ability of the cells to detoxify reactive oxygen species and contributes to cell death. This response takes several days longer than typical hypersensitive cell death responses, thus allowing the limited pathogen growth and restricted sporulation that is characteristic of the WKS1 partial resistance response to Pst.

Different inoculation techniques viz. inoculation of sets by dipping in spore suspension, bud inoculation with hypodermic syringe, bud wrapping by cotton swab dipped in smut suspension and inoculation of underground bud at the time of tillering (end May) were tested to screen against sugarcane smut in the field as well as in the laboratory conditions. In the field, out of the tested inoculation techniques, the maximum disease incidence (60.63%) was observed when buds were inoculated with hypodermic syringe and minimum (8.55%) in inoculation of underground buds at the time of tillering stage. The result showed that inoculation through mechanical injury significantly increase disease incidence but at the same time also affect the bud germination (15.56%). Significant difference in smut incidence was also observed when the inoculation was carried by teliospores and sporidia separately. Key words: Ustilago scitaminea, inoculation technique, screening, resistance, sugarcane.

A comprehensive germplasm evaluation study of wheat accessions conserved in the Indian National Genebank was conducted to identify sources of rust and spot blotch resistance. Genebank accessions comprising three species of wheat-Triticum aestivum, T. durum and T. dicoccum were screened sequentially at multiple disease hotspots, during the 2011-14 crop seasons, carrying only resistant accessions to the next step of evaluation. Wheat accessions which were found to be resistant in the field were then assayed for seedling resistance and profiled using molecular markers. In the primary evaluation, 19,460 accessions were screened at Wellington (Tamil Nadu), a hotspot for wheat rusts. We identified 4925 accessions to be resistant and these were further evaluated at Gurdaspur (Punjab), a hotspot for stripe rust and at Cooch Behar (West Bengal), a hotspot for spot blotch. The second round evaluation identified 498 accessions potentially resistant to multiple rusts and 868 accessions potentially resistant to spot blotch. Evaluation of rust resistant accessions for seedling resistance against seven virulent pathotypes of three rusts under artificial epiphytotic conditions identified 137 accessions potentially resistant to multiple rusts. Molecular analysis to identify different combinations of genetic loci imparting resistance to leaf rust, stem rust, stripe rust and spot blotch using linked molecular markers, identified 45 wheat accessions containing known resistance genes against all three rusts as well as a QTL for spot blotch resistance. The resistant germplasm accessions, particularly against stripe rust, identified in this study can be excellent potential candidates to be employed for breeding resistance into the background of high yielding wheat cultivars through conventional or molecular breeding approaches, and are expected to contribute toward food security at national and global levels.

Leaf rust (Puccinia triticina Eriks.) is a fungal disease of wheat (Triticum spp.), which causes considerable yield loss. Adult plant resistance (APR) is one of the most sustainable approaches to control leaf rust. In this study, field-testing was carried out across ten different locations, followed by molecular screening, to detect the presence of APR genes, Lr34+, Lr46+, Lr67+ and Lr68 in Indian wheat germplasm. In field screening, 190 wheat accessions were selected from 6,319 accessions based on leaf tip necrosis (LTN), disease severity and the average coefficient of infection. Molecular screening revealed that 73% of the accessions possessed known APR genes either as single or as a combination of two or three genes. The occurrence of increased LTN intensity, decreased leaf rust severity and greater expression of APR genes were more in relatively cooler locations. In 52 lines, although the presence of the APR genes was not detected, it still displayed high levels of resistance. Furthermore, 49 accessions possessing either two or three APR genes were evaluated for stability across locations for grain yield. It emerged that eight accessions had wider adaptability. Resistance based on APR genes, in the background of high yielding cultivars, is expected to provide a high level of race non-specific resistance, which is durable.

Two field experiments were conducted to ascertain the potential of micropropagation technique for faster production of seed cane by using tissue culture plants raised through apical meristem culture in first generation (TC 0) followed by clonal propagation through cane setts in next generation (TC1). About 18, 520 plants, produced from a single shoot through micropropagation, were required at row to row and plant to plant spacing of 90 and 60 cm, respectively as compared to 88 quintal of cane seed in conventional methods for planting in an area of one hectare. Multiplication ratio was 100–150 times using tissue culture plants as compared to 11–12 using conventional cane setts, leading to drastic reduction in seed cane requirement. The TC 1 exhibited superiority over vegetatively propagated conventional crop for millable canes and stalk yield by 17 and 10.4 per cent, respectively. Though the single cane weight and cane diameter (non-significantly) were slightly lesser in TC1 as compared to conventional crop, this did not distress its potential as seed crop. The incidence of Ratoon Stunting Disease (RSD) and Leaf Scald Disease (LSD) was very low in TC 1 crop as compared to conventional crop. The findings established the potential of tissue culture technique for the production of quality seed free of pests and pathogens in the existing varieties and rapid multiplication of newly released varieties for quick adoption by the growers.

In the present investigation, optimal media compositions for enhancing the callus induction and subsequent plant regeneration were worked out in sugarcane (Saccharum spp. complex). The field grown somaclones were characterized for yield and quality contributing traits in comparison to donor variety CoJ 88. The MS + 2,4-D (4.0 mg/l) + kin (0.5 mg/l) + sucrose (30 g/l) was found to be the best for the callus induction among the nine media compositions used. The highest shoot regeneration was 34.65% on MS + BAP (0.5 mg/l) followed by 31.07% on MS + IAA (2.0 mg/l) + kin (0.5 mg/l) and the least 11.80% on MS + BAP (2.0 mg/l). The shoot elongation was optimum on MS + IBA (5.0 mg/l). The rooting was carried on MS + NAA (3.0 mg/l) + IBA (2.0 mg/l) + sucrose (70 g/l). The somaclones, on field evaluation, exhibited huge variability for characters viz., tiller number at 120 days of field transfer, number of millable canes, cane height, number of nodes and internodal length at time of harvest and also for HR Brix at 240 and 300 days after field transfer. In R0 generation, the variations induced for the characters such as stalk height after 120 days, HR Brix after 240 and 300 days of field transfer were negatively skewed with most of somaclones having values higher than the population mean values thereby offering adequate scope for improvement cane and quality. Elite somaclones marked for each trait were reported. Out of 228 somaclones inoculated with Cf 08 pathotype of red rot in R1, three were resistant, four were moderately resistant and 221 somaclones showed variable degree of susceptibility, whereas, to Cf 03 pathotype, fourteen were resistant, nineteen were moderately resistant and 195 showed variable degree of susceptibility. Three somaclones exhibited moderate resistance (MR) to both the pathotypes, whereas, donor variety CoJ 88 showed susceptible reaction. These somaclones will further enhance the germplasm for sugarcane breeding.

A comprehensive germplasm evaluation study of wheat accessions conserved in the Indian National Genebank was conducted to identify sources of rust and spot blotch resistance. Genebank accessions comprising three species of wheat-Triticum aestivum, T. durum and T. dicoccum were screened sequentially at multiple disease hotspots, during the 2011-14 crop seasons, carrying only resistant accessions to the next step of evaluation. Wheat accessions which were found to be resistant in the field were then assayed for seedling resistance and profiled using molecular markers. In the primary evaluation, 19,460 accessions were screened at Wellington (Tamil Nadu), a hotspot for wheat rusts. We identified 4925 accessions to be resistant and these were further evaluated at Gurdaspur (Punjab), a hotspot for stripe rust and at Cooch Behar (West Bengal), a hotspot for spot blotch. The second round evaluation identified 498 accessions potentially resistant to multiple rusts and 868 accessions potentially resistant to spot blotch. Evaluation of rust resistant accessions for seedling resistance against seven virulent pathotypes of three rusts under artificial epiphytotic conditions identified 137 accessions potentially resistant to multiple rusts. Molecular analysis to identify different combinations of genetic loci imparting resistance to leaf rust, stem rust, stripe rust and spot blotch using linked molecular markers, identified 45 wheat accessions containing known resistance genes against all three rusts as well as a QTL for spot blotch resistance. The resistant germplasm accessions, particularly against stripe rust, identified in this study can be excellent potential candidates to be employed for breeding resistance into the background of high yielding wheat cultivars through conventional or molecular breeding approaches, and are expected to contribute toward food security at national and global levels.