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Open Access
DEVELOPMENT OF A DIVERSIFIED SET OF Triticum timopheevi BASED FERTILITY RESTORERS IN WHEAT
(Punjab Agricultural University, Ludhiana, 2018) Harmeet Singh; Srivastava, Puja
The study was based on 2100 backcross progenies derived from crosses of fertility restorers with 9 European winter wheat lines. BC1F6 progenies were evaluated at PAU, Ludhiana in 2017-18 for key traits of fertility restoration and anther extrusion, along with preliminary agronomic performance. On the scale of 1-9 for anther extrusion, about 75% progenies exhibited score of 7 and above as high anther extrusion winter wheat lines were used as parents. The range for fertility restoration varied from 12% - 80% in the backcross derivatives. Mean yield of the diversified restorer set was found to be higher than the R-parents. In the restorer evaluation trial, two top yielding lines were line no. 6 (R-1//AMORAK/R-19) and 78 (RS-534//HURLEY/R-4). A total of 26 restorer lines surpassed the highest yielding R-parent. The back cross derivatives involving Amorak as winter parent proved to be superior, followed by Exotic and Maxyl based restorers. R-1//AMORAK/R-19 came out to be best performing in terms of mean yield as well as fertility restoration, whereas R-1//AMORAK/R-1 and RW-17//EXOTIC/R-1, both proved to be best performing for pollen viability and anther size characters. RS-317//HURLEY/R-4 and RS-534//MAXYL/R-28 displayed superiority in terms of tiller number and thousand grain weight, respectively. In the preliminary evaluation of test hybrids, 11.1% F1s showed significant heterosis over commercial checks. The best hybrid from the restorer set was TH 32 (CMS BWL 3275 X R-4//HURLEY/R-1), exhibiting standard heterosis of 29.5%, 21.5%, 25.4% and 28.5%, over checks HD 2967, HD 3086, PBW 725 and PBW 723 respectively. In F2 populations derived from fully fertile F1s, significant association was observed between fertility restoring Rf8 gene with fertility restoration trait.
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Paddy straw, Growth, Lignocellulolytic, Laccase, Endoglucanase, Exoglucanase , -glucosidase, Pretreatment, Biogas, Delftia sp. PP4_S3, Coriolus versicolor MTCC 138
(Punjab Agricultural University, Ludhiana, 2018) Sandeep Kaur; Dhillon, Satwinder Kaur
Soybean harbours many anti-nutrients unsuitable for human digestive tract, among them “Kunitz Trypsin inhibitor (Kti)” is major anti-nutrient inhibiting activity of digestive proteases. Further, the crop is prone to devastating biotic stress i.e. yellow mosaic virus (YMV) prevalent in northern region of the country. Therefore, the present study was planned to identify YMV resistant soybean genotypes carrying null kti gene and cross was made between SL525 x NRC101 to develop YMV resistant and Kti free soybean genotypes. SL525 is YMV resistant but has Kti gene on the other hand NRC101 susceptible to YMV and lacks Kti gene referred to as ‘null kti’. A total of 45 F4 lines of the cross were screened for YMV resistance using infector row technique in kharif 2017. A total of 125 genotypes were selected and grown in kharif 2017 as F5 generation. Molecular markers (gene specific and SSR markers linked with kti locus) were used to screen plants for presence/absence of Kti allele. A total of 65 genotypes were Kti positive, 50 null kti and 10 genotypes were heterozygote. Phenotyping was done by measuring trypsin inhibitor activity (TIA). Consequently, the genotypes lacking Kti allele recorded reduced levels of TIA. Furthermore, YMV resistance of F5 genotypes was confirmed by molecular markers and phenotypic screening. Analysis of variance of F5 genotypes revealed that genetic variation for the morphological and quality traits was present in genotypes. Trypsin inhibitor activity was observed to be positively correlated with germination, days to maturity, plant height, grain yield and seed weight. Non significant direct effect of TIA was observed on grain yield whereas traits such as germination at 6 months after harvest, number of pods per plant, 100-seed weight, plant height, days to maturity showed significant positive direct effect on grain yield. The comparison of performance of null kti and kti genotypes revealed that null kti genotypes had low germination, delayed maturity, less grains and lower TIA values in comparison to kti genotypes whereas traits such as days to flowering, plant height, pods/plants, seed weight, oil content, protein content, fatty acid composition were unaffected by allele present. A total of 14 null kti genotypes had significantly higher grain yield than SL525 and these genotypes were resistant to yellow mosaic disease.
Open Access
Molecular mapping of drought and heat tolerance traits using interspecific cross in chickpea (Cicer arietinum L.)
(Punjab Agricultural University, Ludhiana, 2018) Kushwah, Ashutosh; Sarvjeet Singh
To understand the complex mechanisms of drought and heat stress tolerance, molecular mapping of RIL population through ddRAD-Seq has been undertaken. In this study, all the traits measured were significantly affected by drought stress environment except harvest index as well as by heat stress environment except hundred seed weight and harvest index. The pooled ANOVA for all the traits measured showed highly significant differences in irrigated, rainfed and latesown conditions for genotypic variance as well as for standard error variance. Correlation coefficient exhibited strong association of traits with yield under drought and heat stress conditions. Multiple linear regression analysis revealed the contribution of traits towards the total variation in yield under drought and heat stress conditions. Path analysis explained that biomass followed by harvest index were the major contributor to grain yield under non-stress as well as drought stress and heat stress environment at both locations (Ludhiana and Faridkot) with pooled condition. Principal component analysis also revealed the strong positive association among the traits. In genotypic analysis, a total of 1365 filtered polymorphic and functional SNPs loci were used for linkage map construction through ddRAD-Seq. The average linkage map distance was 4569.09 cM with an average interval of 6.79 cM between the markers. Five QTLs of days to germination (qdg-1), days to flowering initiation (qdfi-1), days to 50% flowering (qdff-1), days to 100% flowering (qdhf-1) and yield (qyld-1) identified using BLUPs under heat stress condition. A total of twenty eight QTLs for days to flowering initiation (qdfi-1 to qdfi-4), days to 50% flowering (qdff-1 to qdff-4), biomass (qbio-1 to qbio-4), yield (qyld-1 to qyld-3), days to 100% flowering (qdhf-1 to qdhf-2), harvest index (qhi-1 to qhi-2), relative leaf water content (qrlwc-1 to qrlwc-2), root-shoot ratio (qrsr-1 to qrsr-2), root dry weight (qrdw-1 to qrdw-2), hundred seed weight (qhsw-1), membrane permeability index (qmpi-1) and fresh root weight (qfrw-1) identified using BLUPs under drought stress condition. These SNPs loci need to be validated in different genetic background and use them in marker-assisted selection for developing superior varieties with enhanced drought and heat tolerance.
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Introgression of cotton leaf curl disease resistance from Gossypium armourianum to G. hirsutum
(Punjab Agricultural University, Ludhiana, 2018) Bhupinder Singh; Rathore, Pankaj
The present investigation was undertaken to evaluate the performance of interspecific F1 hybrid and backcross derivatives of G. hirsutum and G. armourianum cross for response to cotton leaf curl disease as well as to identify the alien segments in the backcross derivatives. Experimental plant material consisted of G. armourianum Acc. PAU 1, G. hirsutum cv. F 1861 and LH 1556, their F1, BC1F1, BC2F1 derivatives. The interspecific F1hybrid between G. hirsutum and G. armourianum was found to be completely resistant to CLCuD. Two of the four BC1F1 plants were observed to be CLCuD resistant. Out of 84 BC2F1 plants, 43 were observed to be resistant, whereas 41 plants were susceptible to CLCuD. Total genomic DNA of the experimental plant materials was amplified employing virus specific primer pairs. Presence of virus DNA was detected in the American cotton parents as well as CLCuD susceptible backcross derivatives. Of the 100 SSR markers used to detect polymorphism in the parental lines, 52 were polymorphic and were used to identify alien segments in CLCuD resistant BC1F1 Plant 2. Eight markers were amplified in this plant demonstrating introgression from the G. armourianum parent. These primers were used to genotype 84 BC2F1plants. The range of G. armourianum specific alleles in the BC2F1plants was observed to be 38.09 through 75.0%. The introgression of G. armourianum specific chromosomal segments in the BC2F1 plants as revealed by SSR markers was independent of the CLCuD response suggesting that none of the eight primers may be linked to CLCuD resistance. Markers TMB 1271 and NAU 6316 exhibited segregation distortion and were transmitted in higher frequency than expected in the BC2F1 progeny.
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Evaluation of an intra-arboreum recombinant inbred line population
(Punjab Agricultural University, Ludhiana, 2018) Lakhvir Kaur; Pathak, Dharminder
The present investigation was undertaken to evaluate the performance of intra-arboreum recombinant inbred lines (RILs) for various quantitative traits and to unravel molecular diversity among a subset of RILs. These two sets including 62 (Set I) and 34 (Set II) RILs along with parent LD 902 and standard check LD 949 were evaluated in a simple lattice design at three locations (Ludhiana, Bathinda, and Faridkot) during kharif 2016. Mean squares for genotypes, locations, and genotype x environment interaction were significant for all the characters except for genotype mean squares for elongation percentage and location mean squares for fibre strength in Set I and location mean squares for micronaire and fibre strength in Set II. The results suggested existence of substantial genetic variability in the experimental plant material, considerable differences among the locations, and profound effect on the expression of seed cotton yield, its components and fibre quality traits. Three RILs namely 431, 241, and 217 in Set I and RIL 105 in Set II significantly out yielded parent LD 902. RIL 105 of Set II appeared to be promising as it recorded numerically higher seed cotton yield than that of LD 949. It also recorded higher values for some of the yield component traits such as number of bolls plant-1, sympods plant-1, and boll weight. Twenty seven RILs of Set I registered significantly higher boll weight than that of LD 949. Ten RILs of Set I and five RILs of Set II possessed at least 18 g/tex fibre strength. Likewise, many transgressive segregants were identified for various traits. Based on SSR diversity analysis of 64 genotypes, RILs were broadly divided into three main clusters. No specific pattern for grouping of genotypes was observed. Substantial variation as revealed by dissimilarity coefficient (0.03 – 0.46) has been observed among different pairs of desi cotton recombinant inbred lines. Further, 29 pairs of RILs recorded at least 0.4 value of the genetic dissimilarity coefficient. The present study has provided a useful insight into performance of desi cotton RILs for various economic traits over locations as well as shed light on the molecular diversity/similarity among the test genotypes.
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Genetic and molecular assessment of breeding methods and mapping novel QTL in soybean (Glycine max L. Merrill)
(Punjab Agricultural University, Ludhiana, 2018) Khosla, Gaurav; Gill, B. S.
Two hundred and seventy five lines from seven different crosses of soybean derived from four different breeding methods viz. pedigree method (PM), single pod descent with selection (SPDS), single pod descent (SPD) and bulk method (BM) were used to assess the efficiency of breeding methods. Based on different yield contributing traits, bulk method was the best method in terms of mean of all lines over two years followed by pedigree method. When the yield of the highest yielding line from each method within each cross was compared with the best check mean, it was observed that the bulk method had the highest yielding line in three out of seven crosses whereas in four crosses it was ranked second. This performance was closely followed by pedigree method. Molecular assessment of the breeding methods was done based on the dissimilarity coefficient among lines within a breeding method using SSR primers. Based on the dissimilarity coefficients, number of unique lines in each breeding method were calculated. SPD method had the highest per cent of unique lines among all the methods in all the crosses. PM and BM had the lowest number of unique lines. Comparison of lines derived from bi-parental and multi-parental crosses revealed that biparental crosses having distantly related parents generated high yielding lines as compared to the multiparental crosses as well as the biparental crosses with less diverse parents. For mapping for YMD resistance genes and seed weight QTL, F2 plants from the cross SL 958 x AGS 456 were used. Segregation ratios in F2, BC1F1 and BC1F2 populations suggested that YMD resistance is controlled by two genes in inhibitory gene action. A total of 269 SSR markers were used for polymorphic studies and the identified polymorphic primers were used for genotyping studies. Bulk segregant analysis was performed with 114 polymorphic markers. Out of 114 markers, 13 markers showed polymorphism in resistant and susceptible bulks. Nine markers were present on C2 linkage group whereas three were present on D1b linkage group. Three major QTL were identified on LG C2 explaining phenotypic variation of 17, 12 and 27 per cent for YMD resistance. One minor QTL was also identified on LG D1b. For seed weight, one major QTL was identified on LG M with Sat_244 and Satt175 as flanking markers having an estimated 19.0 per cent phenotypic variance. Two minor QTLs were also identified on LG D1b.
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Genetic variation for key enzymes involved in nitrogen uptake in American cotton
(Punjab Agricultural University, Ludhiana, 2018) Manjeet Kaur; Paramjit Singh
The present investigation was undertaken to evaluate genetic variation for nitrogen uptake enzymes and to analyze the impact of varied nitrogen levels (0%, 50% and 100% N of RDF) on seed cotton yield, lint yield, and fibre properties of forty nine Gossypium hirsutum genotypes. The enzymes involved in N uptake (nitrate and nitrite reductase) were assayed for Bathinda, at pre-anthesis and post-anthesis stages for each genotype at each nitrogen levels along with chlorophyll and nitrogen content in leaves. The field evaluation of the plant material was undertaken in partially balanced lattice design (7 x 7) with two replications at Ludhiana and Bathinda. Mean squares for genotypes, locations, nitrogen level and genotype x environment, genotype x nitrogen level, genotype x location x interaction were significant for all the characters except for genotype mean squares for fibre fineness. The results suggested existence of substantial genetic variability in the experimental plant material, considerable differences among the locations, and profound effect on the expression of seed cotton yield, its components and fibre quality traits. Genotypes namely LH 900 and F 846 appeared to be promising as it recorded numerically higher enzyme activity at low nitrogen level (0% and 50% N) as compared to recommended dose of nitrogen. Higher enzyme activity of these genotypes at low N dose probably indicates their higher uptake as well as utilization efficiency over other genotypes It also recorded higher values for the yield as well as fibre component traits such as seed cotton yield plant-1, lint yield plant-1, boll weight, ginning out turn, 2.5% span length, and fibre strength. NR enzyme had positive and significant effect on NIR, N content, chlorophyll content, seed cotton yield and lint yield per plant. NIR enzyme showed positive significant effect on chlorophyll content. N content had positive effect on chlorophyll content and seed cotton yield. F846 and LH900 overall performed well as these genotypes were among the best for enzyme activity as well as for yield traits. These genotypes hold future potential for developing new cultivars with improved NUE.
Open Access
Assessment of genetic diversity of shoot fly-resistant sources and transfer QTLs associated with shoot fly resistance into forage sorghums (Sorghum bicolor L.)
(Punjab Agricultural University, Ludhiana, 2018) Arora, Naveen; Sohu, R.S.
The present study consisted of two different sets of genotypes evaluated for variable reaction to shoot fly under field conditions. The first set consisted of a diverse collection of 53 different germplasm lines and the second set consisted of 30 genotypes including 10 cytoplasmic male sterile lines, along with their respective maintainers and 10 restorer lines. On the basis of morphological evaluation under field conditions, the genotypes varied significantly for all the traits under study. ICSA/B 487, ICSA/B 14037, ICSA/B 467, ICSV 93046 and ICSV 700 exhibited high glossiness, pigmented leaf sheath, lower number of eggs and suffered lesser incidence of deadhearts and exhibited resistant reaction to shoot fly while ICSA/B 458, ICSV 25337 and ICSV 25292 exhibited moderate resistance to the shoot fly damage and ICSV- 25280, 25292, 25303, 25316, BTx 623, SSV 74 and Swarna were susceptible to shoot fly. In the germplasm lines significant variation was found for most of the traits except the leaf sheath pigmentation, number of plants with shoot fly eggs and number of shoot fly eggs per plant. Significant positive and negative correlations were found among the important traits for shoot fly resistance like glossy score, leaf sheath pigmentation, shoot fly deadhearts, total number of eggs, number of plants with eggs and trichome density. The inheritance studies based on Line X Tester analysis, revealed that shoot fly resistance is a complex trait and crosses involving resistant parents yielded resistant lines. The lines ICSA 458, ICSA 467, ICSA 487, ICSA 14024 and ICSA 14037 and testers ICSV 700, ICSV 93046, S-35, ICSV 25292 and ICSV 25337 had significant SCA and GCA effects for the traits important for shoot fly resistance. The estimation of biochemical components, revealed that the genotypes exhibiting moderate (ICSA 84, ICSB 52, ICSB 14024) and high (ICSA 467, and ICSB 487) resistance to shoot fly recorded lesser content of total soluble sugars, proteins, higher content of tannins and there was no significant association found for phenol content. From the HPLC analysis of methanol extracts of leaf samples a total of 25 different peaks with 12 known compounds were recorded. Diversity analysis based on GBS based SNP markers grouped the germplasm and parental lines into three major clusters consisting of resistant, moderately resistant and susceptible genotypes. Population structure was analysed using principle component analysis. The first three PC were sufficient enough to explain maximum diversity within population structure. For Genome Wide Association Studies, first three PCs were used as a covariate for which, significant association of SNPs were found for glossy score on chromosome 3, 9, 10 and for percent plants with shoot fly eggs on chromosome 2. For leaf sheath pigmentation, SNPs on all the chromosomes were significantly associated. Similarly, multiple SNPs were associated for percent tiller with shoot fly deadhearts on chromosomes 2,3,4,6,7 and 9. SNPs for plant vigour score were significantly associated with chromosomes 1,2,3 and 6.
Open Access
Marker assisted development of wheat lines with enhanced nutritional quality
(Punjab Agricultural University, Ludhiana, 2018) Kuldeep Kaur; Sohu, V. S.
The aim of present study was the genetic enhancement of grain micronutrient and yellow pigment content in high yielding wheat lines. The study was based on BC1F2 and BC2F2 introgression lines developed from crossing advanced backcross lines of T. durum with grain zinc QTL introgressed from T. monococcum and T. boeoticum, with high yielding varieties of North Western Plain Zone i.e. PBW621, HD2967 and WH1105 and two gene pyramided version of PBW343 (carrying rust resistance genes Yr10, Yr15, Lr24, and Lr28) as recurrent parent. A total of 210 BC1F3 and 210 BC2F3 progenies from different cross combinations were evaluated for grain zinc concentration, yellow pigment content and agronomic traits. Eighteen highly promising BC1F3 progenies (>80ppm zinc concentration) and 15 BC2F3 progenies (>75ppm zinc concentration) were identified and advanced at Keylong as ear to progeny rows. A total of 42 BC1F5 and 43 BC2F5 derivatives were evaluated in a square lattice design and analysis of variance revealed significant variation among the progenies for grain zinc, yellow pigment as well as agronomic and quality traits studied. The BC1F5 progenies from recipient parent WH1105 and tetraploid donor BF13 were recorded with highest grain zinc (64.1-67.9ppm) and iron concentration (29.8-36.0ppm). The highest grain yellow pigment content (5.35 ppm) was observed for the cross involving PBW703 as recipient and BF 21 as tetraploid donor parent. The markers for the previously mapped grain zinc QTL which was identified at diploid level in the donors, were found to be non polymorphic. Monitoring of backcross derivatives with the marker YP7A-2 linked to yellow pigment gene Psy-A1, revealed nine lines positive for high yellow pigment allele (Psy-A1c) indicating introgression from T. durum. To test the effect of environment on grain zinc concentration, eight high zinc BC1F5 lines and ten BC2F5 lines were evaluated in a randomized block design at two locations i.e. Ludhiana and Gurdaspur. The pooled analysis of variation revealed location effect to be significant for grain iron and zinc concentration. Furthermore, BF22/PBW698//PBW698 derivatives were crossed with BWL3560 (carrying Gpc-B1) with the aim of conglomerating enhanced protein content, zinc and yellow pigment content within the same wheat background. A total of 1068 F2 single plants were phenotypically analysed for grain iron and zinc concentration. High zinc lines were shortlisted and 192 F3 lines were sown in square lattice design. Molecular marker analysis of F3 progenies for Gpc-B1 gene (Xucw108) revealed 129 F3 lines to be homozygous positive. Correlation studies revealed that the improvement for four nutritional quality traits (grain protein, zinc, iron and yellow pigment content) was constrained and yield/yield components might have to be compromised.

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