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2013 - 201920
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Subject: Agricultural Biotechnology × End date: 2019 × Start date: 2013 × Type: Thesis × Thesis Type: M.Sc ×
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Now showing 1 - 9 of 20
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    ThesisItemRestricted
    Mapping of Ascochyta blight resistance gene(s)/QTLs from exotic Cicer arietinum L. germplasm in cultivated kabuli chickpea
    (Punjab Agricultural University, Ludhiana, 2019) Lovepreet Kaur; Ajinder Kaur
    Chickpea is an important pulse crop in the world after dry beans. Ascochyta blight caused by Ascochyta rabiei is a major fungal disease of chickpea and can cause upto 100% yield losses under favourable conditions. It is both air-borne and seed-borne disease that spreads rapidly to all the aerial parts including leaves, petioles, flowers, pods, branches and stems leading to rapid collapse and death of plants. The present investigation was undertaken for mapping of Ascochyta blight resistance gene using an interspecific population. The chickpea cultivar, L 552, besides having several agronomically important characters is susceptible to Ascochyta blight and the exotic accession, FLIP 05-43, is resistant to Ascochyta blight. An F2 interspecific population was developed from cross between L 552 (female parent) and FLIP 05-43 (donar parent). Inheritance studies in F2 and F2:3 populations revealed that inheritance of Ascochyta blight resistance was controlled by monogenic recessive gene, that was designated as arr6. A total of 300 SSR markers were used for polymorphism survey of the parents. A considerably low polymorphism of 15.30 % was found between the parents, and 46 polymorphic markers were used for genotyping of F2 population. For generating linkage map, these 46 SSRs were subjected to linkage analysis using MAPDISTO (1.7.6.5) software at LOD of 3, only 31 markers were mapped generating a linkage map of 377.14 cM with eight linkage groups. Using this linkage map, arr6 gene was mapped onto linkage group 4 at a distance of 8.6 cM distal to CGMM072 marker and from NCPGR247 marker, it was located at a distance 16.1 cM. There is a need to identify more SSR markers in the region lying between the markers CGMM072 and NCPGR247 in order to minimize the distance from the arr6 gene and its transfer to the elite cultivar (L 552) for providing durable resistance against Ascochyta blight. Thus, this study provides further prospective for fine mapping and map based cloning of the arr6 gene.
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    ThesisItemOpen Access
    Genetic mapping of brown plant hopper resistance from Oryza rufipogon (Griff.)
    (Punjab Agricultural University, Ludhiana, 2019) Pavneet Kaur; Kumari Neelam
    Brown planthopper (BPH) Nilaparvata lugens (Stal) is one of the most serious insect-pest of rice accounting for the loss up to 52% of annual production. Under high infestation, complete drying and plant death occurs causing “hopper burn”. The primary strategy for controlling BPH is the application of chemical insecticides, which is already proven detrimental not only to the environment, but also to human health. On this account host-plant resistance serves as an important strategy to reduce the damage caused by BPH. Out of 1003 wild species germplasm accessions screened against BPH, O. rufipogon acc. CR100441 has shown consistent resistance reaction (score 1-3) during 3 years of screening under greenhouse conditions. The F1 plant was obtained by making a cross of O. rufipogon acc.CR100441 and PR122. The BC2F2 plants were generated by further back crossing with PR122 and selfing then after. The BC2F2 plants were screened by the standard seedbox technique at seedling stage against BPH biotype 4. The BC2F2 plants were scored phenotypically on a 0-9 scale according to the Standard Evaluation System (IRRI, 1996) as 1-3 resistance, 5 moderate resistance, and 7-9 susceptible plant. From 276 BC2F2 plants, 66 plants were found to be resistant (1-3 score) and 210 plants (5, 7 and 9 score) were susceptible which fitted in 3:1 ratio (susceptible: resistant) indicating the recessive nature of the BPH resistance gene. Genotyping using polymorphic microsatellite markers between PR122 and O.rufipogon acc.CR100441 spanning all the 12 chromosomes of rice was done. The QTL governing BPH resistance was identified on the short arm of chromosome 4 between marker interval RM551 and RM6659. The RM16335 was peak marker, with LOD score of 10.8 and 35.23% contributing towards the phenotypic variation. This information can be used for further fine mapping and cloning of the gene. The pre-breeding BC2F4 lines with BPH resistance can be further utilize in breeding programmes aimed in development of resistant cultivars.
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    ThesisItemOpen Access
    Fine mapping of QTL on chromosome 9 for drought tolerance in maize
    (Punjab Agricultural University, Ludhiana, 2019) Ramandeep Kaur; Sharma, Priti
    Drought is considered as one of the major limiting factors in sustainable maize production all over the world as it causes yield reduction by an average of 15% to 20%. Maize is generally grown in Kharif season but spring maize is now coming up in India. However, the water requirement is very high but farmers are reluctant to see the long term effect of maize cultivation during spring season. To meet the growing demand of water during spring season, enhancement of maize yield can be achieved by developing water efficient maize hybrids. The objective of the study was to identify and to transfer QTL associated with drought tolerance into spring maize inbreds through marker assisted backcross breeding (MABB). A total of 135 F8 recombinant inbred lines (RILs) from the cross between CM123 as the susceptible (female) parent and CM140 as the tolerant (male) parent along with parents were evaluated under control and drought stress conditions for two consecutive seasons. The QTL on chromosome 1, 3, 4, 6, 7 and 9 were identified for drought tolerance under both stress and control conditions. The present study focused on fine mapping of QTL for number of kernel per ear (qKPE) present on chromosome 9 (bnlg1401-umc1634) explaining phenotypic variance of 23.14% under stressed environment. This region was narrow down by designing 50 new SSR markers between the bracketed QTL (qKPE). Seventeen SSR markers showed the polymorphism between CM123 and CM140. These markers along-with previous mapped markers were employed on RIL population. The QTL analysis narrowed down the genetic distance to 3.8 cM from 11.5 cM and physical distance to 691 kb from earlier distance of 15 Mb flanked by two new SSR markers viz. PAU_1143 and PAU_1137. The qKPE is also introgressed through MABB into two spring maize inbreds LM23 and LM24 of hybrid PMH10 for water use efficiency. The foreground selection has been carried out in two generations i.e., BC1F1 and BC2F1. Also, Background selection has been done in BC1F1 to check the background recovery of recurrent parent. The plants carrying the QTL with highest recurrent parent background recovery were selected and again backcrossed to respective parent for generation of BC3F1 population. The BC3F1 plants have been raised during Kharif 2019. The development of drought tolerant PMH10 hybrid will lead to overcome frequent irrigations during spring season and helps to conserve ground water depletion.
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    ThesisItemRestricted
    Linkage map construction in guava F1 population of Allahabad Safeda x Arka Kiran using molecular markers
    (Punjab Agricultural University, Ludhiana, 2018) Jindal, Manish; Mittal, Amandeep
    Guava is a perennial fruit tree grown in tropical and sub-tropical regions of the world. As of now, there are about 160 cultivars available in India. Crop improvement work attempted in India has resulted in release of several superior selections or hybrids. However, the maximum area under guava cultivation is occupied by Allahabad Safeda. Being a cross-pollinated tree (25.7 to 41.3 % cross pollination) guava has a heterozygous genome. Molecular mapping can help us to find the relative positions of the markers as well as the markers co-segregating with the trait of interest that could finally be transferred to the cultivated species. In the present study we have attempted Linkage map construction in guava. A cross between white fleshed Allahabad Safeda and colored fleshed Arka Kiran was attempted in Fruit Science, PAU. We genotyped Allahabad Safeda and Arka Kiran using 167 genomic SSR, 22 EST based and 5 apple color specific markers. Forty eight markers showed polymorphism out of 194 total markers. Polymorphic markers applied on a population of 73 F1 individuals showed segregation. Pattern of marker segregation in the population was scored and analysed with software, MAPDISTO version 1.7.7.0.1.1 (XL 2007) and a genetic linkage map was constructed using stringency parameters of LOD score and recombination frequency set to 3.0 and 0.35, respectively. Out of 48 polymorphic markers, thirty markers were mapped on different linkage groups of guava genome and 6 linkage groups were obtained. The genetic linkage map covered a total of 538.68 cM of the guava genome. Fruits were not set on 2 year old F1 trees so color segregation was studied on leaves as a proxy for fruit color trait. Color in young and mature leaves was measured using miniature leaf spectrometer. The data recorded in terms of anthocyanin reflective index 1 (ARI1) was analysed with the help of mapping software. Two markers mPgCIR93 and mPgCIR21 were mapped to linkage group 2 on positions 54.3 cM and 10.3 cM for leaf color traits at young and mature stage.
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    ThesisItemOpen Access
    Marker assisted mobilization of cry1Ac gene from transgenic chickpea into cultivated chickpea (Cicer arietinum L.) for pod borer [Helicoverpa armigera (Hübner)] resistance
    (Punjab Agricultural University, Ludhiana, 2019) Sharma, Urvashi; Ajinder Kaur
    Helicoverpa armigera (Hübner) poses a serious threat to chickpea production world-wide. cry1Ac gene is known to be most effective in controlling infestation of pod borer. The present investigation was undertaken to incorporate cry1Ac gene from T5 transgenic chickpea lines BS 100B-T5 and BS 100E-T5 into cultivated chickpea (Cicer arietinum L.) varieties PBG 7 and L 552, respectively through crossing and recurrent backcrossing. The BC1F1 progenies [F1 (PBG 7 x BS 100B-T5) x PBG 7] and [F1 (L 552 x BS 100E-T5) x L 552] were tested for cry1Ac gene presence using gene specific primers. ELISA on the positive plants revealed Cry1Ac protein content between 10.57 to 11.72 µg/g leaf tissue. Therefore, on the basis of foreground selection and ELISA, positive BC1F1 plants were propagated to obtain BC1F2 populations, which were also subjected to foreground selection. A BC2F2 population of 83 plants was also grown by selfing [F1 (PBG 7 x BS 100E-T5) x PBG 7] x PBG 7. Foreground selection using cry1Ac specific primers on BC2F2 population resulted in ten (12.04 %) cry1Ac positive plants (1, 2, 8, 9, 12, 20, 26, 33, 39 and 44); their background selection using 12 polymorphic SSR markers revealed average recurrent parent genome restoration of 88.9 %. As a result, BC2F3 population comprising 128 plants was obtained from ten cry1Ac positive BC2F2 plants. Each BC2F3 row of positive plants was then tested using cry1Ac specific primers to identify BC2F2 plants homozygous for cry1Ac gene. The analysis revealed that three (30 %) BC2F2 plants 26, 39 and 44 were homozygous for cry1Ac gene, whereas remaining seven plants 1, 2, 8, 9, 12, 20 and 33 segregated in ratios of 6:1, 2.3:1, 1.25:1, 3:1, 1.6:1, 4:1 and 3:1, respectively.
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    ThesisItemOpen Access
    RNA interference mediated gene silencing in fruit fly [Bactrocera dorsalis (Hendel)]
    (Punjab Agricultural University, Ludhiana, 2019) Mohan. G; Mohanpuria, Prashant
    The oriental fruit fly, Bactrocera dorsalis (Hendel) is serious pest infesting important fruits and vegetables throughout the world. In Punjab, it infests damage of 70-80 per cent each in peach and Kinnow, 60-80 per cent in pear, 50 per cent in jamun, 30-40 per cent each in loquat, fig, plum and mango, and 100 per cent in guava fruits during rainy season and thus, fruit fly is a major concern in Punjab. Control of fruit fly is highly challenging because of its polyphagous nature and unexposed developmental stages and limitations of existing methods. This has now necessitated the biotechnological intervention for fruit fly management. In present investigation double-stranded RNA (dsRNA)-mediated gene silencing in guava fruit fly was attempted. Specific regions of four genes of fruit fly i.e. ecr (ecdysone receptor), rpl19 (a ribosomal protein L19), noa (fatty acid elongase) and v-ATPase-D important for its growth, development, molting, metamorphosis and energy production were cloned in dsRNA expression vector and transformation was done into HT115 (DE3) Escherichia coli strain for dsRNA production. Silencing effects were observed by feeding artificial diet mixed with bacteria expressing dsRNA corresponding to ecr and rpl19 genes to maggot and adult fruit fly. The results showed severe deformities in pupae and adult fruit fly and mortality indicating the feasibility of dsRNA delivery method and silencing of these potential genes of fruit fly. In future, transgenic fruit crops expressing dsRNA of these potential genes of fruit fly and/or spray based dsRNA formulations could be developed for fruit fly management.
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    ThesisItemOpen Access
    Molecular mapping of high grain number QTLs in aromatic rice line RNR-2354
    (Punjab Agricultural University, Ludhiana, 2018) Meelu, Manpreet; Kumari Neelam
    Basmati, the unique quality aromatic rice, is nature‟s gift to the Indian sub-continent. Basmati rice is harmonious combination of aroma, kernel expansion, fluffiness and palatability. Compared to indica varieties, Basmati rice is low yielding primarily because of lower number of grains (70 to 80) per panicle. Therefore, present study is planned to increase the grain number of Punjab Basmati 3 (PB-3). It is a semi-dwarf variety having resistance to the bacterial blight disease of rice (xa 13 and Xa 21 genes), possesses extra- long slender and translucent grains with strong aroma and good cooking and eating quality characteristics. RNR-2354, one of the rice aromatic rice line was found having very high grain number (150- 200) per plant while evaluating and characterizing 400 aromatic rice germplasm lines from all over India. With an objective of improving grain number of PB-3, a cross between PB-3 and RNR-2354 was done to generate F1, F2 and F3 progenies. Phenotypic data were recorded for grain number, grain length and grain width in F2 population and F3 progenies. A total of 5 QTLs designated as qGN-1, qGN-2.1, qGN-2.2, qGN-3 and qGN-5 were mapped on chromosome 1, 2, 3 and 5 using QTL Cartographer mapping software. This information could be further used for narrowing down and dissecting the regions harboring putative QTLs.
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    ThesisItemOpen Access
    Molecular mapping of Fusarium wilt resistance in muskmelon (Cucumis melo L.)
    (Punjab Agricultural University, Ludhiana, 2018) Jaideep Kaur; Sarao, Navraj Kaur
    Muskmelon is an important vegetable crop of the Cucurbitaceae family grown in temperate and tropical regions of the world. Fusarium wilt is one of the major constraints for melon production around the world. Yield losses were reported as high as 100%. It primarily infects the roots resulting in blockage of uptake of water and nutrients, thus killing the plant. Various resistance genes (Fom-1, Fom-2, Fom-3 and Fom-4) specific for specific races have been reported so far. In order to readdress the production bottlenecks in melon within reasonable time frame, deployment of genomic tools such as marker assisted breeding is urgently required. Therefore, this study is frame worked to map the fusarium wilt resistance gene using SSR markers. Two genetically diverse parents “Punjab Sunehri” and “KP4HM15” were crossed and F1 was selfed to generate a F2 mapping population of 154 plants. Phenotypic evaluation on F2:3 families indicated that resistance to fusarium wilt is controlled by single dominant gene, which might be of race 0, 1 or 0, 2. For molecular analysis, a total of 527 simple sequence repeat (SSR) primer pairs were screened for parental polymorphism following bulk segregant analysis (BSA). Linkage analysis indicated that four SSR markers CMCTN35, DM0096, CSWCTT02 and ECM181 were linked to the Fom gene. Marker DM0096 was closest to the gene at 17.6cM distance and the marker CSWCTT02 was mapped at 22.7cM distance from the gene. The four markers were mapped on melon chromosome 5. There is a need to design more flanking SSR markers between DM0096 and CSWCT02 region in order to minimize the distance from the Fom gene and its transfer into the elite cultivar (Punjab Sunehri) for providing durable resistance against fusarium wilt disease. The markers provide further perspectives for fine mapping and map based cloning of the Fom gene.
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    ThesisItemOpen Access
    INTROGRESSION OF crtRB1 AND LcyE GENES FOR HIGH β-carotene INTO QUALITY PROTEIN MAIZE (QPM)
    (Punjab Agricultural University, Ludhiana, 2018) Amandeep Kaur; Malhotra, Pawan Kumar
    Maize (Zea mays L.) being the queen of cereals deficient in Vitamin A which causes the malnutrition and major health problems. Quality protein maize has enhanced level of the amino acids, lysine, and tryptophan over normal maize varieties. However, QPM varieties are low in provitamin A, a precursor of vitamin A which can lead to vitamin A deficiency in human. In the present investigation, the grain quality of QPM inbred is further enriched for β-carotene by introgressing of crtRB1 and LcyE gene through marker assisted backcross breeding. Rare natural genetic variation of crtRB1 and LcyE gene enhances β-carotene in the kernel by blocking its conversion to further components. Traditional yellow maize though contain high kernel carotenoids, but the concentration of provitamin A is quite low (<1.5µg/g) as compared to the recommended level (15µg/g). Development of biofortified maize enriched in provitamin A, lysine and tryptophan thus holds significant potential in the alleviation of micronutrients. Marker assisted stacking of crtRB1, LcyE and o2 were undertaken in the genetic background of QLM13, inbred of PMH1 hybrid. Foreground selection was carried out using gene-specific primers on BC2F1 population of QLM 13 and background selection was carried out using SSR markers to check the recovery of recurrent parent genome. The plants of favorable alleles (crtRB1 and LcyE) and 88.5-90.1% recurrent parent genome recovery were selected and selfed to generate BC2F2 population. Foreground selection was carried out on BC2F2 population using crtRB1 and LcyE gene-specific markers and plant carrying favorable homozygous allele were selfed to generate BC2F3 progenies. Quality analysis for determination of β-carotene and tryptophan analysis was carried out on BC2F3 progenies. The introgressed BC2F3 progenies possessed a high concentration of provitamin A (1.29–11.75 µg/g) as compared to recurrent parent QLM13 (4.69 µg/g). The selected lines of high beta-carotene and tryptophan content were crossed to reconstitute PMH1 hybrid. Introgressed inbred having contrast for pigmentation in glume base and silk with respective to recurrent parents possess great utility for registration and unambiguous identification in the field.