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Chaudhary Sarwan Kumar Himachal Pradesh Agriculture University, Palampur
Himachal Pradesh Krishi Vishvavidyalaya (renamed as Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya in June, 2001) was established on 1st November, 1978.The College of Agriculture (established in May, 1966) formed the nucleus of the new farm University. It is ICAR accredited and ISO 9001:2015 certified institution. The Indian Council of Agricultural Research has ranked this University at eleventh place among all farm universities of the country.
The University has been given the mandate for making provision for imparting education in agriculture and other allied branches of learning, furthering the advancement of learning and prosecution of research and undertaking extension of such sciences, especially to the rural people of Himachal Pradesh.
Over the years, this University has contributed significantly in transforming the farm scenario of Himachal Pradesh. It has developed human resources, varieties and technologies and transferred these to farming community enabling the State to receive the “Krishikarman award” of Govt. of India four times in row for food grain production among small states of the country. Today, the State has earned its name for hill agricultural diversification and the farming community has imposed its faith in the University.
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ThesisItem Open Access Elucidating the role of MYC2 transcription factor in Cucumber mosaic virus infection(palampur, 2019-07-30) Eshwar, Jadhav; Hallan, VipinCucumber mosaic virus (CMV) has the broadest host range among known plant viruses, infecting more than 1,200 species of plants from monocotyledons to dicotyledons. As the effects of CMV is very diverse in nature causing epidemics in diverse crops, it is important to understand the host- pathogen interactions in order to develop plants which can be resistant to plant virus infections. The expression of stress-responsive genes is intimately dependant on its transcriptional control which directly leads to plant response to various stresses. The MYC2 class of transcription factors has recently emerged as a master regulator of the signalling pathways of jasmonic acid (JA) and those of other phytohormones such as abscisic acid (ABA), gibberellins (GAs), and auxin (IAA). Jasmonic acid is the plant hormone that regulates the plant growth, development and defense responses. The MYC2-3 mutant line of Arabidopsis thaliana was used for the analysis of the role MYC2 transcription factor in CMV virus infection. Characterization of MYC2-3 mutant line through jasmonic acid and abscisic acid treatments revealed that the wild type showed reduced root length as compare to MYC2-3 mutant. It was also revealed that the MYC2-3 mutant line has resistance to CMV virus as compared to the wild type plant. Also, SDS PAGE and MALDI-TOF analysis showed that the expression of enzymes and proteins involved in the photosynthetic pathway to be directly affected in the MYC2-3 mutant line on CMV infection. This study could be directly used for development of CMV resistant plant varieties through genome editing in futureThesisItem Open Access Studies on chickpea (Cicer arietinum L.) defence to blight pathogen Ascochyta rabiei(palampur, 2019-07-30) Gaikwad, Dinesh Subhash; Sharma, Kamal DevChickpea (Cicer arietinum L.), a self-pollinated diploid (2n=2x=16) with haploid genome size of ∼740 Mbps, is an important food legume of subtropical and tropical regions and is believed to be originated in the Mediterranean region. In cool and humid regions of the world, chickpea blight caused by Ascochyta rabiei causes huge yield losses. Some accession of C. arientinum and wild Cicer species possess resistance to the disease, however molecular mechanism governing host resistance are poorly understood. With the aim to understand regulation of expression of genes involved in defence to A. rabiei, the expression of six defence genes [Phenylalanine ammonia lyase (PAL), Isoflavone reductase (IFR) Flavanone 3- hydroxylase (F3H), Pathogenesis related protein - 2b (PR-2b), Basic leucine zipper 24 (bzip24) and SNAKIN-2 (Antimicrobial peptides)] was elucidated in resistant and susceptible genotype of chickpea after inoculation (2 h, 12 h, 24 h, 36 h, 72 h) with A. rabiei. Gene expression was studied using quantitative Real-Time PCR and the expression was normalized with two reference genes, Clathrin adaptor complexes (CAC) and ATP-binding cassette transporter (ABCT). The sequences of the mRNAs were retrieved from National Center for Biotechnology Information database followed by identification of coding sequences (CDS) and design of primers. The genes (PAL, IFR, F3H) of the phenylpropanoid pathway that govern phytoalexin production expressed within 2 h after landing of the spores on chickpea surface and expression was considerably higher in resistant HC-1 then susceptible GPF2. The genes for pathogenesis related protein (PR-2b), antimicrobial peptide (SNAKIN-2) and transcription factor (bZIP24) had peaks at 24 h in the resistant genotype indicating maximum expression in resistant genotype at the time of host penetration and subsequent pathogen spread. The susceptible host (GPF2), on the other hand was slow in its response as is evident from low or delayed overexpression as compared to resistant host. It appears that defence to Ascochyta blight in chickpea is a precisely coordinated reaction of the host, where phytoalexin accumulation appears to occur within hours of inoculation, whereas antimicrobial peptides accumulate at the time of host invasion and pathogen spreadThesisItem Open Access Identification of QTLs linked to early maturity and yield-related traits in horsegram (Macrotyloma uniflorum).(CSK HPKV, Palampur, 2019-07-23) Katoch, Megha; Chahota, R.K.Macrotyloma uniflorum is an important, self pollinated diploid (2n=2x=20) food legume with probable genome size of 400Mbps. Limited genomic resources and lack of genetic variation are major constrains in its genetic improvement. Further, horsegram production is hampered due to twining growth habit, longer days to maturity, photosensitivity and indeterminate growth habit. The present study was aimed to construct linkage map of an intraspecific F8 RILs population of 162 individuals derived from HPKM249×HPK4 of horsegram and identification of genomic regions linked to early maturity and yield related traits. Two thousand and eleven molecular markers were screened for parental polymorphism and 493 (25.42 %) were found to be polymorphic among the parents. Of these, 295 were mapped on ten linkage groups at LOD 3.5 spanning 1541.7 cM with an average marker density of 5.20 cM. Analysis of variance of 162 RILs revealed significant differences for all the measured traits. Phenotypic data from the RILs were used to identify QTLs for early maturity and yield related traits by composite interval mapping (CIM). A total of 27 QTLs (LOD ≥ 2.5) were detected across the three environments (Palampur 2016, Palampur 2017, Bajaura 2017) and combined data) for 24 traits. Among these, 15 were major QTLs with PVE greater than ten per cent and five were stable QTLs across locations and years. Phenotypic variation explained (PVE) by QTLs ranged from 6.4 to 53.4 per cent. The highest phenotypic variation (53.4 %) was explained by the QTLs for root length. In conclusion, it is envisaged that the present linkage map, fortified with 295 SSR markers and 27 QTLs for early maturity and yield-related traits would provide genomics tools to breeders for further genetic enhancement of this crop species. Thus, the current study will serve as a strong foundation for further validation and fine mapping of QTLs for utilization in horsegram breeding programs.ThesisItem Open Access REGULATION OF CARBOHYDRATE METABOLISM IN CHICKPEA (Cicer arietinum L.) UNDER DROUGHT STRESS.(CSK HPKV, Palampur, 2019-09-30) Mondal, Kinjal; Sharma, Kamal DevChickpea (Cicer arietinum L.), a cool season annual self-pollinated diploid (2n=2x=16) legume crop with genome size of ∼740 Mbp, is believed to be originated in South-East Turkey and contributes maximum production among the pulses under Mediterranean type environment. Although, chickpea is known for its better drought tolerance than other legumes, terminal drought hinders its production severely even sometimes lead to total crop failures. With the aim to understand regulation of carbohydrate metabolism in chickpea under drought stress, changes in amount of carbohydrate contents, expression of carbohydrate metabolism genes and relative water content (RWC) in leaves of both drought susceptible (ICC 3776) and drought tolerant (ICC 8950) genotypes were studied. Seven genes (ADP-glucose pyrophosphorylase, Phospho-glucan/water dikinase, Iso-Amylase3, Starch synthase4, Beta-amylase1, Beta-amylase3, and Granule-bound starch synthase2) involved in starch metabolism and three genes (Sucrose-phosphate synthase2 like, UDP-glucose pyrophosphorylase, and Cell wall invertase3LX2) involved in sucrose metabolism were studied. Decreased RWC in leaves of the susceptible genotype compared to the tolerant one indicated susceptibility of ICC 3776 to drought. Amounts of total sugars, reducing sugars, non-reducing sugars and sucrose were significantly increased in both the genotypes under induced drought, but in each case, increment was higher in the susceptible genotype. Starch content increased in the susceptible genotype whereas it remained static in the tolerant one. Downregulation of major starch degrading enzyme Beta-amylase3 and simultaneous upregulation of starch synthesizing enzyme starch synthase4 contributed in starch increment despite the upregulation of beta-amylase1 and iso-Aamylase3 in the susceptible genotype. In ICC 8950, starch synthesizing as well as starch degradation enzymes with the exception of Phospho glucan/water dikinase downregulated. Increased accumulation of sucrose content in susceptible genotype compared to tolerant one under induced drought can be attributed to lower expression of Cell wall invertase3LX2 in the susceptible genotype. The study provided insights into sugar metabolism under drought tolerance and drought susceptibility.ThesisItem Open Access ELUCIDATING THE ROLE OF MYC2 TRANSCRIPTION FACTOR IN Cucumber mosaic virus INFECTION.(CSKHPKV, Palampur, 2019-07) Eshwar, Jadhav; Hallan, VipinCucumber mosaic virus (CMV) has the broadest host range among known plant viruses, infecting more than 1,200 species of plants from monocotyledons to dicotyledons. As the effects of CMV is very diverse in nature causing epidemics in diverse crops, it is important to understand the host- pathogen interactions in order to develop plants which can be resistant to plant virus infections. The expression of stress-responsive genes is intimately dependant on its transcriptional control which directly leads to plant response to various stresses. The MYC2 class of transcription factors has recently emerged as a master regulator of the signalling pathways of jasmonic acid (JA) and those of other phytohormones such as abscisic acid (ABA), gibberellins (GAs), and auxin (IAA). Jasmonic acid is the plant hormone that regulates the plant growth, development and defense responses. The MYC2-3 mutant line of Arabidopsis thaliana was used for the analysis of the role MYC2 transcription factor in CMV virus infection. Characterization of MYC2-3 mutant line through jasmonic acid and abscisic acid treatments revealed that the wild type showed reduced root length as compare to MYC2-3 mutant. It was also revealed that the MYC2-3 mutant line has resistance to CMV virus as compared to the wild type plant. Also, SDS PAGE and MALDI-TOF analysis showed that the expression of enzymes and proteins involved in the photosynthetic pathway to be directly affected in the MYC2-3 mutant line on CMV infection. This study could be directly used for development of CMV resistant plant varieties through genome editing in future.ThesisItem Open Access STUDIES ON CHICKPEA (Cicer arietinum L.) DEFENCE TO BLIGHT PATHOGEN ASCOCHYTA RABIEI.(CSKHPKV, Palampur, 2019-07) Gaikwad, Dinesh Subhash; Sharma, Kamal DevChickpea (Cicer arietinum L.), a self-pollinated diploid (2n=2x=16) with haploid genome size of ∼740 Mbps, is an important food legume of subtropical and tropical regions and is believed to be originated in the Mediterranean region. In cool and humid regions of the world, chickpea blight caused by Ascochyta rabiei causes huge yield losses. Some accession of C. arientinum and wild Cicer species possess resistance to the disease, however molecular mechanism governing host resistance are poorly understood. With the aim to understand regulation of expression of genes involved in defence to A. rabiei, the expression of six defence genes [Phenylalanine ammonia lyase (PAL), Isoflavone reductase (IFR) Flavanone 3- hydroxylase (F3H), Pathogenesis related protein - 2b (PR-2b), Basic leucine zipper 24 (bzip24) and SNAKIN-2 (Antimicrobial peptides)] was elucidated in resistant and susceptible genotype of chickpea after inoculation (2 h, 12 h, 24 h, 36 h, 72 h) with A. rabiei. Gene expression was studied using quantitative Real-Time PCR and the expression was normalized with two reference genes, Clathrin adaptor complexes (CAC) and ATP-binding cassette transporter (ABCT). The sequences of the mRNAs were retrieved from National Center for Biotechnology Information database followed by identification of coding sequences (CDS) and design of primers. The genes (PAL, IFR, F3H) of the phenylpropanoid pathway that govern phytoalexin production expressed within 2 h after landing of the spores on chickpea surface and expression was considerably higher in resistant HC-1 then susceptible GPF2. The genes for pathogenesis related protein (PR-2b), antimicrobial peptide (SNAKIN-2) and transcription factor (bZIP24) had peaks at 24 h in the resistant genotype indicating maximum expression in resistant genotype at the time of host penetration and subsequent pathogen spread. The susceptible host (GPF2), on the other hand was slow in its response as is evident from low or delayed overexpression as compared to resistant host. It appears that defence to Ascochyta blight in chickpea is a precisely coordinated reaction of the host, where phytoalexin accumulation appears to occur within hours of inoculation, whereas antimicrobial peptides accumulate at the time of host invasion and pathogen spread.ThesisItem Open Access EVALUATION AND UTILIZATION OF SSR MARKERS FOR GENETIC DIVERSITY CHARACTERIZATION IN SEABUCKTHORN.(CSKHPKV, Palampur, 2019-07) Dangat, Smitesh Sunil; Sharma, Ram KumarSeabucktthorn (Hiippophae rhamnoiides,, diiplloiid 2n=24) iis a diioceous,, wiind polllliinatted,, tthorny shrub bellongs tto famiilly Ellaeagnaceae.. In Indiia,, iitt grows natturalllly iin hiigh allttiittude and colld-ariid regiions of Ladakh,, Siikkiim,, Jammu & Kashmiir,, Hiimachall Pradesh,, and Uttttrakhand.. Itt iis economiicalllly and ecollogiicalllly iimporttantt mediiciinall pllantt due tto iitts hiigh iimmense mediiciinall properttiies butt tthere iis unavaiillabiilliitty of superiior variietty// sellecttiion avaiillablle for llarge scalle commerciiall cullttiivattiion iin Indiia.. Furtthermore,, valliidatted miicrosattelllliitte marker resources are allso lliimiitted iin tthiis pllantt speciies.. Therefore,, a ttottall of 882 novell SSR markers were miined from iin-house H.. rhamnoiides ttranscriipttome datta.. Among allll,, abundance of ttrii-repeatts (49%) follllowed by dii-repeatts (46%) was observed.. Furtther,, evalluattiion and uttiilliizattiion of pollymorphiic SSR makers for genettiic diiversiitty assessmentt of 245 accessiions of Seabucktthorn (H.. rhamnoiides) dettectted hiigh average allllelle frequency (5..45 allllelles// priimer) and hiigh hetterozygosiitty (Ho:: 0..77;; He:: 0..75).. Neiighbor-Joiiniing dendrogram,, priinciipall coordiinatte anallysiis (PCoA) and Bayesiian modell based popullattiion sttructture anallysiis grouped allll tthe 245 iindiiviidualls iintto ttwo majjor groups wiitth hiigh wiitthiin popullattiion gene fllow suggesttiing tthatt tthere are ttwo genettiic poolls operattiing iin tthe natturall seabucktthorn popullattiions of ttrans--Hiimallayan Leh and Keyllong regiions of Indiia.. The resulltts of tthiis sttudy wiillll be usefull for iidenttiifiicattiion and sellecttiion of pottenttiiall genottypes tto expediitte tthe conservattiion and genettiic iimprovementt programme iin seabucktthorn.. Genome-wiide miicrosattelllliitte marker resource creatted duriing tthiis sttudy can be valliidatted and uttiilliized for futturiisttiic evolluttiionary and variious genottypiic sttudiies iin seabucktthorn.ThesisItem Open Access IDENTIFICATION OF GENOMIC REGIONS ASSOCIATED WITH PLANT ARCHITECTURAL TRAITS IN LENTIL (Lens culinaris Medik)(CSKHPKV, Palampur, 2019-07) Kamble, Ravindra Siddheshwar; Chahota, R.K.Lenttiill ((Lens culliinariis Mediik subsp.. culliinariis)) iis an auttogamous diiplloiid ((2n=2x=14)),, cooll season ffood llegume crrop cullttiivatted glloballlly.. Genomiic rresourrces iin llenttiill arre lliimiitted iin comparriison tto ottherr ffood llegumes,, prriimarriilly due tto llarrge genome siize and llack off genettiic varriiattiion.. Furrttherr,, llenttiill prroducttiion iis hamperred due tto grrowiing underr shorrtterr day llengtth and narrrrow genettiic bases off rrelleased cullttiivarrs rresponsiiblle fforr yiielld llosses iin llenttiill.. Pllantt heiightt,, numberr off brranches,, lleaff arrea,, rrespecttiive posiittiion off tthe orrgans ((lleaff,, sttem,, ffllowerrs and ffrruiitts)),, arre genettiicalllly conttrrolllled,, butt allso depend on enviirronmenttall iinfflluences orr human iintterrventtiions.. The prresentt iinvesttiigattiions aiimed tto consttrructt iinttrraspeciiffiic llenttiill lliinked map usiing 91 RIILs popullattiion tto iidenttiiffy rregiion lliinked tto pllantt arrchiittectturrall ttrraiitts.. Siix hundrred fforrtty tthrree SSR marrkerrs ((iinclludiing prreviiouslly publliished anchorr marrkerrs)) werre scrreened fforr parrenttall pollymorrphiism and 343 ((27..90 %)) werre ffound tto be pollymorrphiic among tthe parrentts.. 334 pollymorrphiic marrkerrs,, werre mapped on seven lliinkage grroups att LOD 4..0 spanniing 987..0 cM wiitth an averrage marrkerr densiitty off 2..95 cM.. The rresulltts rrevealled 80 new SSRs assiigniined on tthe lliinkage map.. Anallysiis off varriiance rrevealled siigniiffiicantt diifffferrences fforr allll tthe 8 measurred arrchiittectturrall ttrraiitts bettween „„L830‟‟ and „„Prrecoz‟‟.. The ANOVA off 91 RIILs rrevealled siigniiffiicantt diifffferrences fforr allmostt allll tthe ttrraiitts.. Phenottypiic datta ffrrom tthe RIILs werre used tto iidenttiiffy QTLs fforr pllantt arrchiittectturrall ttrraiitts by composiitte iintterrvall mappiing ((CIIM)).. A ttottall off 29 QTLs ((LOD ≥ 3..0)) werre dettectted on seven lliinkage grroups.. Phenottypiic varriiattiion expllaiined ((PVE)) by QTLs rranged ffrrom 11..9 tto 41..9 perr centt.. The hiighestt phenottypiic varriiattiion ((41..9 %)) was expllaiined by tthe QTLs fforr 100--seed weiightt.. IIn concllusiion,, iitt iis enviisaged tthatt tthe prresentt lliinkage map,, fforrttiiffiied wiitth 334 SSR marrkerrs and 29 QTLs fforr pllantt arrchiittectturrall ttrraiitts woulld prroviide genomiics ttoolls fforr ffurrttherr genettiic enhancementt off tthiis crrop speciies.. Thus,, tthe currrrentt sttudy woulld serrve as a sttrrong ffoundattiion fforr ffurrttherr valliidattiion and ffiine mappiing off QTLs fforr uttiilliizattiion iin llenttiill brreediing prrogrrams.ThesisItem Open Access DEVELOPMENT OF In Vitro SYSTEMS OF Colleus forskohlii FOR FORSKOLIN PRODUCTION(CSKHPKV, Palampur, 2019-07-06) Sharma, Mohini; Bhattacharya, AmitaThe roots and tubers of Coleus forskohlii,, an endangered herb of family,, Lamiaceae are medicinally important because of the presence of forskolin,, an alkaloid popular for its anttii-obese,, bllood-pressure-lloweriing,, nerve-regenerattiing and iinttraocullarpressure lloweriing properttiies.. Owiing tto adenyllatte-cycllase-sttiimullantt acttiiviitty,, forskolliin iis used iin ttreattiing gllaucoma,, congesttiive-cardiiomyopatthy and astthma.. Indiiscriimiinatte up-roottiing of tthe pllantt for forskolliin has lled tto acutte scarciitty of raw matteriiall.. There are no cullttiivattiion practtiices and tthe forskolliin llevells are allso llow iin rootts and ttubers.. Therefore,, an objjecttiive of tthe tthesiis was tthe devellopmentt of miicropropagattiion prottocolls for UP NBRC-10,, UP NBRC-13,, J&K,, Tamiillnadu,, Punjjab and Haryana accessiions of C.. fforskohlliiii.. Resulltts revealled maxiimum shoott mullttiiplliicattiion (approx.. 70 shootts) occurred on 0..8% agar gelllled (w//v) fullll sttrengtth MS mediium conttaiiniing 3% sucrose,, 7..50 μM BAP and 2..50 μM IBA butt shoott ellongattiion was maxiimum on 3% sucrose suppllementted fullll sttrengtth PGR-free-MS mediium.. In comparattiive growtth performance Haryana accessiion supportted maxiimum number of rootts,, whiille Tamiillnadu had maxiimum roott llengtth.. In hardeniing experiimentt,, maxiimum surviivall was recorded iin sand duriing summer.. When variious auxiins were used,, maxiimum callllusiing wiitth miiniimum rhiizogenesiis occurred iin lleaf and sttem expllantts of UP NBRC-13 and J&K att 10 μM 2,,4-D.. Adventtiittiious roott formattiion was maxiimum iin lleaves of Haryana accessiion on 2..5μM NAA suppllementted 0..8% agar gelllled and lliiquiid MS mediia.. In tthe second objjecttiive,, tthe forskolliin conttentt iin rootts,, sttem and lleaves of iin viittro and fiielld grown accessiions were measured usiing HPTLC//UHPLC.. Forskolliin was hiighestt iin rootts of iin viittro grown Haryana accessiion.. In case of adventtiittiious rootts,, forskolliin was maxiimum iin Tamiillnadu butt llow iin Hayana accessiion.. Ten ttiimes hiigher iincrease was recorded att 12..5 μM IBA.. The fiindiings pave tthe way for en masse propagattiion of hiigh-forskolliin-yiielldiing C.. fforskohlliiii pllantts for iindusttriiall use.
