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ThesisItem Restricted 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 SinghThe 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.ThesisItem Restricted 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 KumarIn 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.ThesisItem Open Access Imparting root knot nematode resistance through grafting in tomato (Solanum lycopersicum L.)(Punjab Agricultural University, Ludhiana, 2020) Sumeet Singh; Kulbir SinghThe present investigation was conducted at Vegetable Research Farm, Department of Vegetable Science, Punjab Agricultural University, Ludhiana for “Imparting root knot nematode resistance through grafting in tomato (Solanum lycopersicum L.)” during Autumn season of 2015-2016 and 2016-17. The work plan was divided into three experiments based on different grafting methods which further divided into two different sites i.e. infested with root knot nematode and non-infested soil. Ten rootstocks with combination of two different scions (Punjab Sartaj and Punjab Ratta) were used as experiment material. For infested plots pure culture of nematodes were raised by taking galled roots of tomato plants from infested plots than galls were teased and freshly hatched second stage juveniles were raised and used for inoculums. It was observed that plant and fruit characters like plant height, fruit weight, harvest duration, marketable yield as well as total yield increased in grafted plants over nongrafted plants both in nematode free as well as Nematode infested plots. Further it was also revealed that plants grafted with Tube graft method observed highest success rate and was even responsible for earliness as well as high marketable yield in tomato plants. On evaluation of rootstocks plants grafted on EC-119197 recorded highest plant height, higher total and marketable yield, higher fruit weight and prolonged harvest duration both in Nematode infested and nematode free plots. Whereas plants grafted on rootstock PNR-7 recorded early flowering as well as early picking in infested plots. Rootstock L-097 recorded maximum fruit TSS while plants grafted on rootstock EC-119197 recorded maximum pericarp thickness. No significant effect was observed when grafted and non-grafted plants were examined for lycopene content. Overall rootstock EC-119197 can be used as a resistant rootstock against root knot nematode in addition it can fetch higher yield and better fruit quality.ThesisItem Open Access Identification and characterization of salt tolerant genotypes and salt tolerant gene in brinjal (Solanum melongena L.)(Punjab Agricultural University, Ludhiana, 2020) Talwar, Dilpreet; Kulbir SinghThe present investigation was conducted at vegetable research farm, Department of Vegetable Science, Punjab Agricultural University, Ludhiana for “Identification and characterization of salt tolerant genotypes and salt tolerant gene in brinjal (Solanum melongena L.)” during the year 2016-18. The experiment was laid out in split plot design with four salinity levels (control, 2 dS/m, 4 dS/m and 6 dS/m) as main factor and 102 genotypes of brinjal as sub main factor. The desired levels of salinity were induced using sodium chloride, magnesium sulphate and calcium chloride in ratio 2:1:1 in field as well as in pot study. These genotypes were categorised as 19 genotypes under tolerant, 39 as semi-tolerant and 44 were susceptible to salinity on the basis of nursery and morphological screening trials. Tolerant genotypes were further evaluated for physico-chemicals characters. It has been observed from the investigation that plant growth and fruit characters of brinjal i.e. plant height, leaf area, root fresh and root dry weight; number of fruits per plant, fruit yield decreased as the salinity level increased. The results showed that brinjal genotypes SL-8-PB-1-3-1-4 followed by KBSR-343-1 and BL-215 performed better upto 4 dS/m levels of salinity with a slight decline in yield traits, while at 6 dS/m yield reduction was high, which was 12%, 28% and 28.5% respectively as compared to control. The total chlorophyll and carotenoid pigments of brinjal leaves decreased as the salinity level increased. Whereas, proline, protein, phenols and MDA content increased as the salinity level increased with maximum increase in SL-8-PB-1-3-1-4 genotype. Brinjal genotype SL-8-PB-1-3-1-4, KBSR-343-1 and BL-215 performed better for germination and survival rate at all salinity levels. By using NCBI database, SOS-3 gene sequence for brinjal was identified. Designed primer was amplified and the resulted PCR product was sent to Europhin Company for identification of haplotypes in brinjal. In the present investigation, SL-8-PB-1-3-1-4 (Small Long), KBSR-343-1 (Small round) and BL-215 (Long) emerged as ideal genotypes for growing under high salinity in field as well as pot conditions. Quality of these genotypes found least affected under salt stress. Thus, three brinjal genotypes (SL-8-PB-1-3-1-4 (Small Long), KBSR-343-1 (Small round) and BL-215 (Long)) may be recommended for future studies in salt affected area of Punjab.ThesisItem Open Access Assessment of genetic diversity in carrot genotypes using morphological, biochemical and molecular markers(Punjab Agricultural University, Ludhiana, 2018) Davinder Singh; Dhillon, Tarsem SinghThe study entitled “Assessment of genetic diversity in carrot genotypes using morphological, biochemical and molecular markers” was carried out at Vegetable Research Farm, Department of Vegetable Science, Punjab Agricultural University, Ludhiana. Eighty one genotypes of carrot were characterized by morphological, biochemical and molecular markers during 2015-16 and 2016-17. Analysis of variance showed significant differences for 19 quantitative traits with respect to treatment, year and treatment x year. Maximum total root yield was obtained from genotypes viz. PC-161, PC-15, PC-43 and PC-103 (red), PCO-30 and PCO-5 (orange) and PCP-2 and PCP-1 (purple). On the basis of morphological diversity, 81 genotypes were distributed into ten clusters. Cluster X was the best for root weight and anthocyanin content, cluster III for days to 1st root harvest and root girth, while dry matter content, total sugar content and carotene content were maximum in cluster V. The highest intra cluster distance has been observed from the cluster VII (162.9), while inter cluster distance was maximum between II and cluster X (43678.5). The cumulative proportion of variation explained by the four PC-axes was 85.12 %. Seventeen polymorphic SSR markers differentiated genotypes into three groups where cluster I consisted of 48 genotypes followed by cluster II with 28 genotypes and cluster III had 5 genotypes. The morpho-biochemical and SSR markers were effective tools to identify closeness among genotypes, so future strategy involves utilizing more SSR markers culled from various linkage mapping could be effectively and efficiently used to fasten the breeding programs and to design trait specific carrot breeding.ThesisItem Restricted Mitigation of cold stress in tomato (Solanum lycopersicum L.) through physiological interventions(Punjab Agricultural University, Ludhiana, 2017) Meena, Yogendra Kumar; Khurana, Daljit SinghTo study ameliorate effect of phenolic compounds against low temperature stress in tomato under open field conditions during the winter season of 2014-15 and 2015-16. The cultivar Punjab Ratta comprised the plant material were given foliar application twice, first 15 days after transplanting followed by another spray after a fortnight of different concentrations of phenolic compounds viz., salicylic acid, sulfosalicylic acid, methyl salicylic acid, benzoic acid and acetyl salicylic acid (0.1mM, 0.5mM & 1.0mM) in order to evaluate their effect on morphological, physiological, yield and quality traits. The observations recorded during two years revealed that low temperature significantly reduced the growth biomarkers (plant height, number of branches, number of leaves, shoot and root length, and total biomass of plant, leaf are index, specific leaf area, leaf weight ratio, relative growth rate and net assimilation rate), physiological and biochemicals attributes viz., relative water content, photosynthetic pigments (chlorophyll and carotenoid content), ascorbate peroxidase activity, yield attributes (number of flowers per cluster, number of fruits, fruit weight and fruit yield) and quality parameters (total soluble solid, lycopene, β-carotene and α-tocopherol). On the other hand, low temperature significantly enhanced the specific leaf weight, electrolyte leakage, accumulation of osmoprotectants (proline, total soluble sugar and sucrose), catalase, peroxidise, hydrogen peroxide and lipid peroxidation. However, application of phenolic compounds mitigated cold stress by increasing abovesaid growth, physiological and biochemicals, yield and quality attributes and furthermore by accumulating osmoprotectants, increasing the activity of antioxidant enzymes and reduced the specific leaf weight, electrolyte leakage, hydrogen peroxide and lipid peroxidation. Among all treatments, salicylic acid @ 1.0mM was found to be the most effective to mitigate low temperature stress in tomato.ThesisItem Open Access Optimizing soil matric potential and climate based irrigation scheduling to potato under different establishment methods(PAU, 2015) Ahuja, Sanjeev; Khurana, D.S.An Investigation under experiment-I was conducted with the objective to determine the most appropriate installation depth of tensiometer with suitable soil matric potential for higher tuber yield and water use efficiency of potato under furrow-ridge irrigation system at Vegetable Research Farm, Punjab Agricultural University, Ludhiana during 2011-12 and 2012-13. Three levels of soil matric suction (S); 20, 35 & 50 kPa at three installation depths (D) of tensiometer; 20, 30 & 40 cm and a control (traditional practice) were replicated thrice in RCBD. The pooled analysis of data showed that treatments S20D20 and S35D20 recorded 86 and 82% higher marketable tuber yield compared to the control. The WUE under S35D20 treatment was upto177% higher than control when data was pooled over two years. Highest dry tuber yield was recorded in S35D20 treatment among all the treatments during both the years. The analysis of pooled data for marketable and graded tuber yield, average tuber weight, discarded percent of tubers, root dry weight, lower content of reducing sugars and higher percent of starch and protein content, plant height at harvest and above ground dry matter accumulation 30, 60 DAP and at harvest also favoured S35D20 and S20D20 treatments. However averaged over two years, WUE and WUEi of S35D20 was more than double as compared to control treatment along with higher marketable tuber yield as 49 % irrigation water was saved. The results revealed that treatment S35D20 was the best treatment for irrigation scheduling in the region. Further, experiment-II was conducted in split plot design with four replications to compare the climatic based irrigation scheduling with S35D20 under ridge-furrow (M1) and bed-furrow planting (M2) system during 2012-13 and 2013-14. Two rows were planted on each bed under furrow-bed planting system. The treatments consisted of five irrigation levels: I1; IWCPE (Irrigation water to cumulative pan evaporation) ratio 1.0, I2; IWCPE ratio 1.25, I3; IWCPE ratio 1.5, I4; S35D20 (tensiometer installed at 20 cm soil depth maintaining SMP of -35 kPa) and I5; control (farmers practice in the region). The ridge-furrow method of planting (M1) showed its advantage over the bed-furrow method of planting (M2) in terms of recording higher WUE, average tuber weight, marketable tuber yield, large and medium sized tuber yield, plant height at 60, 75 DAP and at harvest as well as above ground dry weight accumulation at the time of harvest. Almost three times higher water use efficiency was recorded by S35D20 as compared to control treatment (farmer’s practice) under M1 method of planting. This irrigation treatment was superior in terms of plant height, above ground DMA at 60 DAP and at harvest, graded and marketable tuber yield, starch and protein content and has lower discarded percent of tubers among all other treatments. The results of two experiments suggest that scheduling irrigation based on soil matric potential at -35 kPa SMP with tensiometer installing at soil depth of 20 cm under ridge-furrow irrigation system was better option under North-Western plains of India. Adopting this threshold corresponds to 3 to 4 irrigations of 45 to 45.5 mm each depending on the rainfall during potato growing period. Two separate irrigations of 50 mm each were applied, first as pre-planting irrigation and second irrigation to substantiate emergence. Thus, application of single irrigation two weeks after the complete emergence of crop and remaining 2-3 irrigations at 17 days interval may save irrigation water as compared to the traditional irrigation practice.
