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Open Access
Processing suitability of chili varieties for value added products
(Punjab Agricultural University, Ludhiana, 2020) Manpreet Kaur; Sachdev, Poonam A.
The present investigation entitled ―Processing suitability of chili varieties for value-added products‖ was carried out with the objectives to assess the physicochemical, phytochemical and antioxidant activity of Punjab Sindhuri and CH-27 chili varieties, development of chilli paste with enhanced shelf stability, drying characteristics of chili varieties and preparation of value added products with greater shelf stability and to extract oleoresins by using conventional and non-conventional methods. The CH-27 contained highest total soluble solids (5º Brix), total solids (20.7%), ash content (1.47%), total sugars (4.4mg/100 gm), crude fibre (9.81%), total phenolics (276 mg/100ml), ascorbic acid (132.17 mg/100 gm) and capsaicin (1.17%), while Punjab sindhuri contained highest total chlorophyll (10.43), carotenoids (12.05mg/100 gm), β-carotene (5.85mg/100 gm) and lycopene (3.2µg/100 gm). Among different drying techniques, freeze drying at -40ºC showed significantly highest retention of physicochemical, phytochemical and antioxidant properties in both chili varieties. During 8 months storage (room as well as refrigeration), honey chili sauce containing CH-27 (red) showed utmost retention of total phenolics (679.46mg/100ml), β-carotene (308 mg/100 gm), lycopene (4.59µg/100 gm), per cent inhibition of DPPH (52.02%), FRAP (3.43 mM/100 gm), while honey chili sauce containing Punjab sindhuri (red) significantly retained maximum total solids (84.49%) ascorbic acid (5.54mg/100 gm), color a value (4.84), ash content (2.86%) and total sugars (63.56 gm/100 gm). Red and green form of both chili varieties were processed into the paste and stored at the room as well as refrigeration temperature. The chili paste yield was found significantly higher for Punjab sindhuri (green and red) than CH-27. Treatment containing 0.2gm KMS + 0.2gm sodium benzoate + 1ml acetic acid + 5gm NaCl per 100 gm of chili paste was found to be effective for the better retention of physicochemical, phytochemical and antioxidant activity during 10 months duration of storage. Highest oleoresin yield i.e.7.1% from Punjab sindhuri red variety was observed with ultrasonication method.
Open Access
Inheritance and molecular mapping of cotton leaf curl disease resistance in synthetic amphiploid × Gossypium hirsutum cross
(Punjab Agricultural University, Ludhiana-, 2020) Vij, Suruchi; Pathak, Dharminder
The present investigation was undertaken with the objectives to study inheritance of cotton leaf curl disease and to map the genes imparting CLCuD resistance. For this, the available CLCuD resistant F1 hybrids between a 'Synthetic polyploid' and G. hirsutum Acc. PIL 43 were used for the development of BC1F1 population using PIL 43 as the recurrent parent. CLCuD resistance was dominant over its susceptibility. Two genes were found to govern resistance to CLCuD as revealed by the appearance of resistant and susceptible plants in the ratio of 1:3 in the BC1F1 generation. The CLCuD resistance genes were mapped on chromosome A01 and D07. SNP marker SD07_2729958 was found to be associated with the target gene at a distance of 24.8 cM on chromosome D07. The gene on chromosome A01 was flanked by markers SA01_115554458 and SA01_288632 which were 41.1 cM and 37.3 cM away from the target gene, respectively. The linkage map spanned a length of 12606.89 cM with an average distance of 6.82 cM between adjacent markers. Similar recombination rates were observed in the A- and D-subgenomes. A set of 210 BC1F3 progenies were evaluated along with four checks (PIL 43, LH 2108, F 2228 and F 2164) for yield and fibre traits. Nineteen progenies having tenacity in the range of 31.3 g/tex through 34.7 g/tex were observed to be significantly superior to the recurrent parent (28.8 g/tex). Similarly, progenies outperforming the recurrent parent for fibre length and fineness were also recovered.
Open Access
Weed management in organically grown basmati rice-durum wheat cropping system.
(Punjab Agricultural University, Ludhiana, 2020) Hasam, Hasamuddin; Simerjeet Kaur
The present study on “Weed management in organically grown basmati rice-durum wheat cropping system” was conducted at Students’ Research Farm, Department of Agronomy, Punjab Agricultural University, Ludhiana during Kharif 2017 and 2018 and Rabi 2017-18 and 2018-19. Nine non-chemical weed control treatments were tested along with one conventional agriculture treatment using herbicide metsulfuron + carfentrazone 25 g ha-1 as a post emergence in wheat and pretilachlor 750 g ha-1 as pre emergence in rice crop. Conventional and deep tillage with green manure seed rates at 50, 75 and 100 kg ha-1 plus 25% higher plant density and one hand pulling, and weedy and weed free treatments were tested in puddled basmati rice (Pusa basmati 1121); while conventional tillage and deep tillage with 50% higher plant densities, zero tillage with and without residues, straw mulch at 6 t ha-1 and bed planting plus 25% higher plant density followed by hand pulling, and weed free treatments were studied in durum wheat (WHD 943). The lowest number of seeds of Dactyloctenium aegyptium, Echinochloa colona, Trianthema portulacastrum and Cyperus iria in rice and Anagallis arvensis, Medicago denticulate, Rumex dentatus, Chenopodium album and Phalaris minor in durum wheat was recorded mostly in upper soils (0-15 cm) in conventional tillage whereas in deep tillage most of seeds were displaced to deeper layer (1530 cm) in both basmati rice and durum wheat. Population of E. colona and Eclipta alba were found lower in all weed control treatments as compared to weedy check but grain yield of rice was found statistically at par among all treatments. In durum wheat, weed density and biomass were found lower in bed planting treatment during both years of study as compared to rest of treatments. Different weed control practices comprising of tillage, green manuring, plant density and straw mulching have non-significant effect on quality of basmati and durum wheat. The maximum grain yields were obtained from weedfree and bed planting with 25% more plant density followed by hand pulling.
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Molecular mapping and interspecific hybridization for hull-less seed trait in pumpkin for value addition in food processing industry
(Punjab Agricultural University, Ludhiana-, 2020) Barinder Kaur; Dhatt, Ajmer Singh
The present investigation entitled ‘Molecular mapping and interspecific hybridization for hull-less seed trait in pumpkin for value addition in food processing industry’ was conducted in the Department of Vegetable Science, Punjab Agricultural University, Ludhiana from 2016 to 2020. Segregating populations were developed using hulled and hull-less genotypes of Cucurbita pepo for understanding the genetics and molecular mapping of hull-less seed trait. Chi-square analysis of segregating populations revealed that single recessive gene is responsible for controlling this trait. The sequence based bulked segregant analysis of two extreme bulks (hulled and hull-less) of cross HP111 × HLP36 along with their parental lines identified one QTL on chromosome 12 (Cp4.1LG12) between 1891261 to 3863267 bp covering 1.97 Mb genomic region with 396 number of SNPs for hull-less seed trait. In interspecific hybridization, total 916 crosses between C. pepo (HLP36, HLP44, HLP53, HLP72) and C. moschata (HM1404, HM108, HM1343, HM1022, HM2211, HM6711) were attempted to transfer hull-less seed trait, however success was achieved in 5.57% crosses only. The fruit setting, number of seeds/fruit, seed germination and survival of F1 seedlings were high when C. pepo was used as female parent in cross HLP36 × HM1343 (66.67%, 15.00, 45.24% and 49.48%), followed by HLP36 × HM1022 (22.73%, 23.00, 44.35%, 47.06%) and HLP44 × HM1022 (13.79%, 12.00, 45.85%, 45.46%), respectively. However, in reciprocal crosses (C. moschata × C. pepo) success rate was very poor, which was overcome through in vitro ovule culture after 17 to 19 days of pollination in cross HM1343 × HLP53 and HM6711 × HLP72. The hybridity of putative Cucurbita interspecific hybrids was confirmed through molecular, morphological and micromorphological markers and their results were found to be highly consistent. The phenotyping of F2 populations of interspecific crosses confirmed the transfer of hull-less seed trait. Findings of the present study will facilitate in genetic dissection of candidate gene(s) and to enhance productivity, improve quality and multi-season cultivation of hull-seeded pumpkin in India.
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Marker assisted breeding for introgression of genetic male sterile ms10 gene from hot pepper to heat tolerant bell pepper
(Punjab Agricultural University, Ludhiana-, 2020) Menisha Rani; Jindal, Salesh Kumar
In the present investigation, marker assisted selection (MAS) strategy was employed to introgress genetic male sterile (GMS) ms10 gene from the donor parent MS-12 (Capsicum annuum var. annuum), popular GMS line, into the genetic background of eight heat tolerant bell pepper (C. annuum var. grossum) inbreds, to develop new GMS lines in bell pepper. The chi-square test (χ2) results of six F2 and BC1F2 populations showed that the ratio well fitted to the Mendelian ratio i.e. 3:1, validated the non-discriminated segregation of the male sterility (MS) trait possessing ms10 gene. The analysis confirmed that the MS trait is inherited by a single recessive gene. Further, the recently identified molecular marker ‘AVRDC-PP12’ linked to the ms10 gene was employed to test the segregation in backcross generations of bell pepper incorporated with the ms10 gene. A total of 576 and 360 individuals in BC1F1 and BC2F1, respectively, 72 plants of each BC1 and 60 plants of each BC2 were screened (foreground selection) for the ms10 gene by using SSR marker. Among the eight backcrossed populations screened, six were showed compatibility with the ms10 gene. The results were analyzed by χ2-tests to know whether the observed number of plants in each class fitted well to the anticipated genetic ratio of 1:1. The molecular marker in each backcrossed series displayed a good fit in-to the expected ratio of Mendelian segregation i.e. 1:1 for one gene in backcross population(s), which was non-significant at a probability level of 0.05, indicating the relationship with pepper male sterility ms10 gene. Simultaneously, targeted gene positive (Ms10ms10) non-pungent (pun1) heterozygous plant(s) were identified in BC2F1 generations using BF7 and BF9-SCAR marker. A total of 27 BC2F1 Ms10ms10 plants of MSSM-2 population were segregated in a 5: 22 non-pungent to pungent ratio, and 26 Ms10ms10 plants of MSSM-3 segregated in 3 (non-pungent): 23 (pungent) ratio. The rest of the four BC2F1 populations, MSSM-1, MSSM-9, MSSM-17 and MSSM-21 have pungent heterozygous male fertile (Ms10ms10 Pun1pun1) plant. The background recovery rate of the recurrent parent phenome (RPP) in BC2F1 progeny was estimated by measuring the shift in means for horticultural traits. Based on a paired 't' test analysis, out of six, the MSSM-1 and MSSM-2 generation was shown to be non-significant for all 19 studied traits followed by MSSM-3 (18), indicating a successful recovery of the recipient genome through marker-assisted backcross breeding after successful introgression of the ms10 gene. Finally, from this work six gene positive (ms10) GMS lines namely, MSSM-1, MSSM-2, MSSM-3, MSSM-9, MSSM-17 and MSSM-21 were obtained. The newly developed GMS lines will provide great economic value for non-pungent hybrid bell pepper seed production in the near future.
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Agrobacterium mediated genetic transformation of pigeon pea (Cajanus cajan L. Millsp.) for resistance to spotted pod borer Maruca vitrata
(Punjab Agricultural University, Ludhiana, 2020) Manjinder Singh; Ajinder Kaur
The present investigation dealing with introduction of cry1Ab gene for resistance to Maruca vitrata into pigeonpea (Cajanus cajan L. Millsp.) through Agrobacterium-mediated in planta transformation was carried out using three pigeonpea genotypes i.e. AL 15, AL 201 and PAU 881. Two methods of in planta transformation were used for generating transgenic plants, including inoculation of pricked embryo axes and floral dip transformation. Out of seven experiments on in planta transformation method involving inoculation of pricked embryo axes, only two (OD600 of Agrobacterium broth = 0.6 - 0.7 containing 100 mM acetosyringone, 1 h dipping of 2-day old pricked seedlings) gave positive results. A total of 5,022 half seeds were treated that gave rise to 4059 plants, out of which 15 (representing AL 15 and AL 201 genotypes) were PCR-positive with an overall transformation frequency of 0.36 %. Out of 15 primary transformants, only 11 plants showed transcript accumulation by semi-quantitative RT-PCR. The primary transformants were also analyzed for chimerism by ELISA-based protein accumulation in selected branches. Out of a total of 182 branches of 10 RT – PCR positive plants analyzed, 57, 43 and 49 branches had Cry1Ab protein content equal to or more than 1.08 µg/g [Positive calibrator, (PC) 5.0 ppb], 0.58 µg/g (PC, 2.5 ppb) and 0.08 µg/g (PC, 0.5 ppb), respectively. Besides, there were 33 such branches which did not show accumulation of Cry protein indicating the presence of chimerism in primary transformants. The putative transgenics were advanced to T1 generation, out of 904 plants analyzed, 97 were PCR-positive, thus exhibiting transformation efficiency of 11.70 % (with genotype AL 201) and 10.24 % (with genotype AL 15). Twenty three T1 plants were randomly taken for determination of Cry1Ab protein content, out of which 12 showed high protein content of more than 0.72 µg/g. These plants showed positive results in PCR and RT-PCR, and further used for determining the efficacy of cry1Ab gene against second instar larva of Maruca vitrata using insect bioassay. Both flowers and pods of these 12 T1 pigeonpea plants were used for bioassay experiments. All 12 T1 plants showed restricted increase in larval weight as compared to nontransgenic (control) plant. No insect mortality was observed, but larvae fed on two transgenic plants i.e. 201-344 and 15-537 showed no adult emergence as compared to other transgenic and control plants where adult emergence was normally observed. Both of these two plants (201-344 and 15-537) had high Cry protein content (0.88 µg/g) as compared to other transgenic plants, which established clearcut positive correlation between amount of protein accumulated and inhibition of M. vitrata larval growth. Further, T1 plants were advanced to T2 generation that were maintained in transgenic glasshouse. In case of floral dip transformation, a total of 139 flower buds were treated and 454 seeds were obtained. Ten out of 454 putative T1 plants were observed to be PCR-positive with a transformation frequency of 2.2 %.
Open Access
Influence of insecticides on growth and development of Bemisia tabaci (Gennadius) infesting cotton
(Punjab Agricultural University, Ludhiana, 2020) Simranjit Kaur; Vijay Kumar
The study on ‘Influence of insecticides on growth and development of Bemisia tabaci (Gennadius) infesting cotton’ was undertaken on 50 days old plants of Bt cotton cultivar, RCH 773 under screen house conditions at Entomological Research Farm, Department of Entomology, PAU, Ludhiana during crop season 2018 and 2019. Effect of repeated application of different insecticides (flonicamid 50 WG @ 100 and 200 g/ha, pyriproxifen 10 EC @ 625 and 1250 ml/ha, spiromesifen 22.9 SC @ 250 and 500 ml/ha, fipronil 5 SC @ 1000 and 2000 ml/ha, ethion 50 EC @ 1000 and 2000 ml/ha and bifenthrin 10 EC @ 450 and 900 ml/ha) on the biology and oviposition preference of B. tabaci; biochemical constituents of cotton leaf and relative expression of vit and jhe gene in B. tabaci were recorded. The results revealed that fecundity of whitefly was significantly lower in spiromesifen 22.9 SC @ 500 ml/ha (18.93 and 29.22 eggs/female) and higher in bifenthrin 10 EC @ 900 ml/ha (43.47 and 62.00 eggs/female) and fipronil 5 SC @ 2000 ml/ha (38.99 and 47.04 eggs/female) over untreated control (32.49 and 40.64 eggs/female) during 2018 and 2019, respectively. Pyriproxifen and spiromesifen significantly prolonged the incubation period, total nymphal duration, pupal period and total developmental period and had minimum egg, nymphal and immature stages’ survival of whitefly. Similarly, pyriproxifen and spiromesifen recorded minimum adult longevity, nymphal growth index, growth index of immature stages and resurgence ratio of B. tabaci. Bifenthrin neither prolonged the duration of various developmental stages nor affected nymphal growth index of B. tabaci. However, sex ratio (female:male) of B. tabaci was increased in fipronil (2.18 and 2.03) and bifenthrin (2.06 and 2.00) over untreated control (1.62 and 1.41) during 2018 and 2019, respectively. Spiromesifen (23.89 and 40.33 eggs/leaf) treated plants were least preferred for oviposition by B. tabaci whereas bifenthrin and fipronil (128.78, 250.56, and 123.33, 219.33 eggs/leaf, resp.) treated plants were most preferred over untreated control (81.00 and 120.33 eggs/leaf) during 2018 and 2019, respectively. The results obtained from biochemical studies revealed that bifenthrin and fipronil treated leaves showed increase in total soluble sugar, reducing sugar, total soluble protein, free amino acid and crude protein content of cotton leaf and decrease in starch content over untreated control. Bifenthrin also decreased the total phenol and tannin content of cotton leaves. pH, total soluble sugars, reducing sugars, sucrose and free amino acids had significant positive correlation with fecundity, adult longevity and oviposition preference of whitefly whereas, phenols and tannins had negative correlation. Bifenthrin and fipronil did not show a significant effect on the relative expression of jhe and vit gene in B. tabaci. Overall results confirmed the resurgence of B. tabaci in Bt cotton with repeated application of bifenthrin and fipronil.
Open Access
Mapping and transfer of GENES/QTLs for nematode (Meloidogyne graminicola) resistance from Oryza glaberrima into Oryza sativa L.
(Punjab Agricultural University, Ludhiana, 2020) Gurwinder Kaur; Vikal, Yogesh
Rice root knot nematode (Meloidogyne graminicola) is one of the emerging constraints for rice production, causes about 50% yield losses in field conditions and 98% yield losses in pot experiments. The identification and exploitation of resistant rice genotypes is one of the economic and sustainable option to minimize the yield losses caused by M. graminicola infestations. The present investigation was undertaken to identify the QTLs associated with rice root knot nematode resistance as well as the transfer of resistance to elite rice cultivars. A total of 42 accessions of O. glaberrima along with O. sativa cultivar PR121 were screened in triplicates during kharif 2016 and kharif 2017 in nematode infested sick plot at initial nematode population density of one juvenile per gram of soil. Reproduction factor of rice root knot nematode among O. glaberrima accessions was found to be <1 while in O. sativa cultivar PR121, reproduction factor was >1. Out of 42 accessions, three accessions (IRGC102196, IRGC102538 and IRGC102557) were found to be highly resistant, thirty-three accessions were resistant, and six accessions were moderately resistant. Data on different morphological traits (plant height, root length, fresh shoot weight, fresh root weight, dry shoot weight and dry root weight) revealed that no statistically significant differences were found among O. glaberrima accessions whereas PR121 exhibited significant reduction in all growth parameters in nematode infested sick plot as compared to controlled conditions. The BC1F1 population derived from the cross of O. glaberrima acc. IRGC102206 × PR121 was used for mapping of rice root knot nematode resistance. The BC1F1 plants were screened against M. graminicola in triplicates and genotyped using 84 polymorphic SSR markers. Both phenotypic data and genotypic data was analyzed and a total of 13 QTLs associated with gall number, gall index and different morphological traits were mapped on chromosomes 1, 3, 4, 5, 6 and 8. Out of 13 QTLs, three QTLs associated with gall number were identified; two QTLs designated as qGN4.1and qGN4.2 were mapped on chromosome 4 and one QTL, qGN6.1, was mapped on chromosome 6. QTL associated with gall index (qGI6.1) was found to be co-localized with qGN6.1. Similarly, qGN6.1 QTL was co-localized with plant height and root length. Due to less coverage by SSR markers, we simultaneously performed BSA-QTLseq (Bulked segregant analysis coupled with QTL-seq approach) analysis and identified QTLs for nematode resistance on chromosomes 1, 2, 3, 4, 5, 6, 11 and 12. The genomic interval of all QTLs was narrowed down to 1–2 Mb using QTL-seq analysis. The development of SNP based molecular markers from identified QTL regions will further help to saturate the linkage map and to identify closely linked markers to rice root knot nematode resistance. The identified markers will further fasten the improvement of genotypes for rice root knot nematode resistance through marker assisted breeding approach. Based on introgressed genomic regions from O. glaberrima carrying QTLs for nematode resistance the BC1F1 resistant plants were selected and backcrossed to generate BC2F1 and subsequently BC3F1 progenies for transfer of nematode resistance in the background of PR121. The data generated from this study can serve as valuable genomic resources for rice breeding programmes.
Open Access
Identification and characterization of high temperature stress responsive genes in maize (Zea mays L.)
(Punjab Agricultural University, Ludhiana, 2020) Ashok Babadev Jagtap; Vikal, Yogesh
Heat stress due to climate change is an emerging issue for maize breeders as it largely affects the yield. The present study focuses to elucidate molecular mechanisms and underlying genes, or QTLs associated with heat stress tolerance in maize. The transcriptional studies of maize leaves, pollens and ovules using heat stress susceptible and tolerant inbred lines, namely LM11 and CML25, respectively revealed a total of 2,164 (1127 up-regulated and 1037 down-regulated) differentially expressed genes (DEGs) between LM11 (HS) and CML25 (HT) samples, with 1151, 451 and 562 DEGs were identified in comparisons of corresponding leaf, pollen and ovule samples, respectively. Functional annotations of DEGs showed that many of them were related to transcription factors (TFs) viz. AP2, MYB, WRKY, PsbP, bZIP and NAM, heat shock proteins (HSP20, HSP70 and HSP101/ClpB), as well as genes related to photosynthesis (PsaD and PsaN), antioxidation (APX and CAT) and polyamines (Spd and Spm). KEGG pathways analyses explicated that metabolic overview pathway and secondary metabolites biosynthesis pathway, with involvement of 264 and 146 genes, respectively were highly enriched in response to heat stress. A bioinformatics pipeline was used to call and type SNPs from RNA-seq reads and applied it to transcriptomic data of LM11 and CML25. A total of 554,423, 410,698 and 596,868 polymorphic SNPs were identified respectively among leaf, pollen and ovule of the LM11 and CML25. A total of 100 genome-wide SNP based KASP assay markers were developed and validated as well as subsequently genotyped on 90 F2 individuals derived from the cross of LM11 × CML25. The success of SNP conversion rate was 71%. In addition, F2 population and their parental inbreds were genotyped using 94 polymorphic SSR markers. The 175 F2:3 families during late March (Spring 2017) were evaluated for heat stress under field and glass house conditions. Heat stress significantly affected all the morpho-physiological and yield contributing traits. Grain yield was positively associated with ear weight, number of kernels per ear, pollen viability, pollen shedding durations and chlorophyll content. Furthermore, secondary traits like membrane thermostability, days to anthesis and silking, anthesis-silking interval, canopy temperature, leaf firing and tassel blast showed significant negative impact on grain yield in both field and glass house conditions under heat stress. Both genotyping and mean phenotypic data of each component trait was analyzed for single marker analysis (SMA) and composite interval mapping (CIM) using WinQTL Cartographer. A linkage map of 1857.1 cM in total length was constructed by applying both SSR and SNP markers. A total of 11 QTLs were detected for 7 traits on chromosomes 1, 3, 4, 6, 7 and 9 with phenotypic variance ranged from 8.67 to 29.62 per cent. Four of these QTLs, qKPE6.1, qPV6.1, qCC9.2 and qLF4.1, accounted for above 15 per cent of phenotypic variation, and might be considered as major QTLs for heat tolerance. The data generated in present investigation laid the foundation for future work to uncover genes and mechanisms critical for the development of heat-resilient maize using genetic and biotechnological approaches.

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