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Dr. Y. S. Parmar University of Horticulture & Forestry, Solan
Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, was established on 1st December, 1985 with the objective to promote education, research and extension education in the fields of Horticulture, Forestry and allied disciplines. Late Dr. Yashwant Singh Parmar, the first Chief Minister and the architect of Himachal Pradesh perceived the importance of Horticulture and Forestry to develop and improve the State economy which led to the establishment of this University. Its history lies in erstwhile Himachal Agricultural College, Solan, established in 1962 and affiliated to the Panjab University. It became one of the campuses of Agriculture Complex of Himachal Pradesh University on its formation in 1970. Consequent upon the establishment of Himachal Pradesh Krishi Vishvavidyalaya in 1978, this campus became its Horticulture Complex and finally in 1985, assumed the status of a State University, being the only University in the country engaged exclusively in teaching, research and extension in Horticulture and Forestry.
The University is located at Nauni in Solan District of Himachal Pradesh, 13 km from Solan on Solan-Rajgarh Road, at an elevation of 1300 metres above mean sea level. Solan town is situated on national highway (NH-22) and is well connected by train and bus services.
The University has four constituent colleges, out of which, two are located at the main campus Nauni, one for horticulture and the other for forestry, having 9 and 7 departments, respectively. The third College i.e., College of Horticulture & Forestry is located at Neri in Hamirpur District on Nadaun-Hamirpur state highway, about 6 Km from Hamirpur town and is well connected with bus service. The college offers three Undergraduate Degree Programmes i.e. BSc (Hons.) Horticulture, BSc (Hons.) Forestry and B. Tech. Biotechnology and MSc degree programme in a few subjects. The fourth college i.e. College of Horticulture and Forestry, Thunag (Mandi) is located at Thunag District Mandi. This college offer BSc (Hons.) Horticulture and BSc (Hons.) Forestry degree programme. In addition, there are five Regional Research Stations, 12 Satellite Stations and five Krishi Vigyan Kendras (KVKs) situated in different zones of the State.
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ThesisItem Open Access CLONING AND CHARACTERISATION OF SUGAR PARTITIONING AFFECTING (SPA) PROTEIN GENE FROM POTATO (Solanum tuberosum L.) AND ITS EXPRESSION ANALYSIS IN RESPONSE TO HIGH TEMPERATURE(NAUNI,UHF, 2020-11) NEWAR, DIVYANI; ANUPAMA SINGHABSTRACT Potato (Solanum tuberosum L.) holds the status of the world’s most important nongrain food crop and ranks third in terms of total production after rice and wheat. Temperature is the most important factor affecting the growth, development and tuberization in potato. It influences the metabolic and photosynthetic rates thus, affecting the dry matter accumulation. A fundamental process in understanding the plant growth and development is carbohydrate partitioning. One of the proteins found to be affecting the carbohydrate partitioning in plants is Sugar Partitioning Affecting (SPA) protein gene. The present study was carried out to clone and characterize the SPA gene in potato in response to high temperature stress. Full length amplicon of approximately 500 bp size was cloned and sequenced. Expression analysis using semi-quantitative RT-PCR was done under controlled (18oC) and heat stress (26oC) condition in three different tissues namely leaf, root and tuber in four potato varieties namely Kufri Surya (heat tolerant) and Kufri Chandramukhi, Kufri Jyoti and Kufri Pukhraj (heat sensitive). In case of leaf, the expression of SPA gene was highest in Kufri Surya under heat stress conditions as compared to heat sensitive varieties. In case of root and tuber, the expression was highest in Kufri Chandramukhi. Sucrose estimation was done in three potato varieties namely Kufri Surya, Kufri Chandramukhi and Kufri Jyoti in response to high temperature. Kufri Surya was found to have highest sucrose concentration under control (18oC) as well as under heat stress (30oC) conditions at the end of 48hrs and 72hrs treatment as compared to the other two varieties which was found to be statistically significant. Further work on this gene in potato would be of great value in establishing this gene as a candidate for developing heat tolerant potato cultivars.ThesisItem Open Access Agrobacterium MEDIATED IN-PLANTA TRANSFORMATION OF POTATO WITHANNEXIN GENE(NAUNI,UHF, 2020-11) SHARMA, SAMRITY; ANUPAMA SINGHABSTRACT Potato is a starchy tuberous crop that holds and important position among all horticultural crops. However, climatic changes have greatly reduced the overall quality and production of potatoes. It has been reported that damages due to high temperature conditions is the most uncontrolled factor in the potato crop production. Hence it’s very important to develop various efficient methods to induce tolerance in potato varieties to abiotic stress. Conventionally, breeding is being done in various crops to improve a desired trait but it’s not beneficial in case of potatoes so genetic transformation is preferable. In the present study, the potato varieties ‘Kufri Surya’ and ‘Kufri Chandramukhi’ were transformed using annexin gene construct through Agrobacterium-mediated in-planta transformation method which doesn’t involve tissue culture steps. In- silico identification of eleven potato annexin genes was done using Phytozome software. Peptide sequence alignment of potato was checked with peptide sequence Arabidopsis ATANN1 annexin gene, it was done using Clustal Omega software. Confirmation annexin gene construct in the Agrobacterium stock was done by isolating the plasmid DNA followed by restriction digestion with EcoR1 and Sal1. A band of 950bp was observed. In-planta transformation was carried out by incubating sprouted tubers with AB minimal media and a transformation efficiency of 52.17% in ‘Kufri Chandramukhi’ and 34.04% in ‘Kufri Surya’ observed. Confirmation of transformation in potato tubers with the STANN1 annexin gene was done through PCR analysis using NPTII specific primers. Transformed plants were subjected to heat stress conditions at 30°C. Expression analysis of other 5 annexin genes in transformed Kufri Surya tubers was done using Semiquantitative Real time PCR analysis. Quantitative RT-PCR analysis was done to check the expression of Aquaporin and importin proteins in Kufri Surya transformed lines kept under heat stress condition it was observed that aquaporin concentration was fairly higher in all transformed plants whereas importin expression levels were downregulated in all Kufri Surya plants. The difference in the chlorophyll contents of transformed and control Kufri Surya plants kept under heat stress was checked. There were significant differences in the chlorophylla and chlorophyllb contents as compared to total chlorophyllcontent.ThesisItem Open Access BIOMASS ACCUMULATION AND ESTIMATION USING in vitro CALLOGENESIS IN Saussurea costus (Falc.) Lipsch. – AN ENDANGERED MEDICINAL PLANT(NAUNI,UHF, 2020-11) KARTIK; SHARMA, RAJNISHABSTRACT Saussurea costus (Kuth) is an endangered medicinal plant due to its anti-inflammatory, anticancer, anti-ulcer, and hepatoprotective activities. Therefore, the present investigation was carried out to explore any alternative approaches to extract the different bioactive compounds available in this medicinal herb, for which the whole plant is uprooted and due to which it has been listed as endangered in Himalayan region. The effect of different factors (like type of explants, growth regulators, photoperiod and effect of elicitor) affecting callogenesis for biomass accumulation and estimation in S. costus using different explants was evaluated. Among four explants viz., root, leaf, cotyledon and hypocotyl, root explant was found to be best with a maximum average callus induction (83.29%) in minimum average number of days (14.46) when cultured on Murashige and Skoog (MS) medium fortified with 2,4-D (1.0 mg/l) and BAP (0.50 mg/l). The maximum callus induction and biomass yield were observed at 16:8 hours photoperiod. In addition, comparative study of phytoconstituents among in vivo (root and leaf) and in vitro (root callus, leaf callus, cotyledon callus, hypocotyl callus and roots from leaf explants) samples revealed that maximum alkaloid (7.28%), flavonoid (6.41%) and terpenoid (1.61%) contents were found in in vivo root samples followed by in vivo leaf extracts. Likewise, in vitro roots resulted in higher alkaloid (3.81%), flavonoid (2.51%) and terpenoid (0.69%) contents than in vitro calli derived using all the explants. Among the in vitro obtained calli, maximum alkaloid, flavonoid and terpenoid contents were observed in root calli viz., 3.40%, 2.03% and 0.53%, respectively. The phytoconstituent concentrations in in vitro calli derived using root and leaf explants were also enhanced when MS medium was augmented with 150 µM of salicylic acid as elicitor. It was concluded that root explant was found to be best for callus growth, biomass accumulation as well as extraction of phytoconstituent contents followed by leaf among all the explants. Hence, the present study could be beneficial towards scaling up the extraction of these bioactive compounds as well as in conserving this targeted medicinally important endangered herbThesisItem Open Access IN VITRO PROPAGATION OF GINGER (Zingiber officinale Rosc.) CULTIVAR SG-1156(NAUNI,UHF, 2020-11) DHAUTA, SHIVANI; THAKUR, MANISHAABSTRACT In the present investigation, a technique for in vitro propagation of ginger (Zingiber officinale Rosc.) cv. SG-1156 has been developed. Actively growing buds from ginger rhizomes were surface sterilized with 0.1% mercuric chloride (HgCl2) for 11 min, to give maximum number of uncontaminated buds (60%) and bud survival (40%). Maximum in vitro bud establishment of 60 % was achieved on establishment medium E5 comprising of MS salts with 0.5 mg/l BA and 0.1 mg/l NAA. Highest rate of shoot multiplication (1:6) was observed in the medium M1 comprising of MS salts with 0.5 mg/l BA and 0.1 mg/l NAA, which was similar to E5. The shoots formed on M1 were healthy and stout with maximum (6.5 cm) average shoot length and number of leaves.With the increase in number of sub cultures, rate of shoot multiplication, shoot length and per cent rooting increased reaching a maximum after 6thsubculture. Healthy rooting was observed at the base of shoots during shoot multiplication phase only, thus eliminating an additional step of rooting. As the number of subcultures increased average root number and length increased, reaching a maximum of 6.3 roots per shoot after 6th passage with average root length of 7.5 cm. Fully rooted in vitro plantlets were hardened after 2nd subculture till 6th subculture and percentage hardening increased from 60% to 95 % from 1st to 6th subculture. Various potting mixtures were tested for hardening of in vitro regenerated plants and maximum survival (95%) was obtained in potting mixture comprising of sand: soil: FYM (1:1:1). During bio hardening best growth and plantlet survival (80 %) was observed by drenching the soil with 20 ml of jeevamrit (3 %) whereas, drenching with different amount of PGPR led to low survival percentage (31.1%) of plantlets. Hardening was significantly affected by season and maximum hardening success of 95 % was observed in monsoons followed by autumn (50%) and summer (40%).ThesisItem Open Access GENETIC FIDELITY STUDIES ON IN VITRO PROPAGATED GISELA-5 (Prunus cerasus x Prunus canescens) – CHERRY ROOTSTOCK(NAUNI,UHF, 2020-09) DHIMAN, AARZOO; THAKUR, MANISHAABSTRACT Thein vitro cultures of Gisela-5 maintained in the department of Biotechnology were further subcultured in routine at four weeks interval. Three DNA based molecular markers were used to test the genetic fidelity of in vitro shoot cultures, micropropagated hardened plants and donor mother plant of Gisela-5. Out of twenty RAPD primers, fourteen primers amplified the genomic DNA of Gisela-5. A total of 23 bands were scored, out of which 17 were found to be monomorphic. For ISSR analysis, fifteen primers were used, out of which 10 primers were able to amplify the genomic DNA of Gisela-5. A total of 19 bands were amplified with 52% polymorphism. Of all the 36 SCoT primers used, only 16 primers produced amplified bands in all the samples of Gisela-5. Out of 38 amplified bands, 26 bands obtained were monomorphic with 32% polymorphism. The size of the amplified bands ranged between 100-5000 base pairs. The Dendrogram based on UPGMA clustering method was constructed using similarity matrix. The Dendrogram showed 94% similarity coefficient in RAPD, 76% in ISSR and 92% in SCoT thus showing some somaclonal variations in among in vitro shoot cultures and tissue culture raised plants thus recommending the establishment of fresh cultures periodically to obtain true-to-type planting materialThesisItem Open Access EVALUATION OF rol B TRANSFORMED CLONES OF APPLE ROOTSTOCK MERTON 793 FOR ROOTING ABILITY AND ITS RELATION TO GENE EXPRESSION(UHF,NAUNI, 2020-08) THAKUR, SANGAM; MODGIL, MANJUABSTRACT Rooting and growth characteristics of rol B gene transformed and non transformed plants of apple rootstock Merton793 were investigated. Transformed plants included five clones namely, E10(1), E10(2), E16(2), E23(1) and E27(2). In vitro rooting showed that all the transgenic clones rooted to 50-66% in auxin free medium and 66-83% on 0.2 mg/l IBA supplemented medium used for short period, whereas non transformed controls resulted in 50% rooting only on latter medium. The number of roots per rooted shoot in transgenic clones ranged from 2-8 which was more as compared to the control shoots. Growth analysis of potted plants maintained in glasshouse revealed that only a few transgenic plants showed a reduction in increase in stem length and number of nodes as compared to the untransformed controls, while no significant differences were found in internode length. Stem girth of transgenic plants, in particular, E27(2) increased significantly. RT-PCR confirmed the expression of rol B gene, while Real Time PCR analysis resulted in high level of expression in rol B transgenic plants whereas no expression was recorded in nontransformed controls. The level of fold change varied from 2.06-2.56 among the different transgenic lines. High expression of transgene is related to improved rooting ability and stronger root system in Merton793 which is considered as difficult to root conventionally while no correlation was found to growth parameters except some changes in stem girth and length. In future, further studies are needed to evaluate the growth of scion cultivars after grafting on transgenic rootstock plants and to assess the rooting on cuttings under misting.ThesisItem Open Access GENETIC FIDELITY STUDIES ON IN VITRO PROPAGATED PLUM (Prunus salicina L.) CVS. SANTA ROSA AND FRONTIER(UHF,NAUNI, 2020-09) RAKSHANDHA; THAKUR, ManishaABSTRACT The present studies were undertaken to establish clonal fidelity in Plum cvs Santa Rosa and Frontier by taking six year old axenic in vitro cultures and two year old tissue culture raised plants growing in the field as experimental material. Genomic DNA was isolated from fresh leaves of in vitro shoot cultures and plants of Plum cvs. Santa Rosa and Frontier. RAPD, ISSR and SCoT markers were used to amplify DNA of Plum cultivars. Out of 28 RAPD primers used, 18 and 15 primers were able to amplify the genomic DNA of Plum cv. Santa Rosa and Frontier, with the monomorphism of 96.55% and 100%, respectively and a similarity coefficient of 0.84 between both the cultivars. All the 15 ISSR primers were able to amplify the genomic DNA of both the Plum cultivars and produced a monomorphism of 92.72% and 95.45% in Santa Rosa and Frontier respectively. A homology of approximately 65% homology between both the cultivars. Out of 26 SCoT primers, only 10 SCoT primers amplified the genomic DNA of Plum cvs. Santa Rosa and Frontier resulting in monomorphism of 96.77 and 90% in both the cultivars, respectively. On comparing Plum cvs. Santa Rosa and Frontier, 84-97% similarity was observed between the two. All the three molecular markers were able to amplify the genomic DNA of Plum cultivars under study with reproducible results. From the above studies it can be deduced that Plum cvs. Santa Rosa and Frontier have a common lineage due to which large amount of similarity can be seen in both of them at the molecular level. In vitro propagation can be successfully used for clonal multiplication of Plum cvs. Santa Rosa and Frontier without any undesireable variations, as validated using RAPD, ISSR and SCoT primersThesisItem Open Access COST EFFECTIVE IN VITRO PROPAGATION OF GINGER (Zingiber officinale ROSC.) CV. HIMGIRI(UHF,NAUNI, 2020-09) ANITA; THAKUR, ManishaABSTRACT Ginger (Zingiber officinale Rosc.) is a perennial herb belonging to the family Zingiberaceae. In the present studies, cost effective micropropagation protocol of ginger cv. Himgiri has been developed by replacing sucrose and agar with low cost alternatives. 40 g/l table sugar and 15 g/l isabgol proved to be the best as a carbon source and gelling agent. Out of different combination tried, isabgol and table sugar showed maximum multiplication rate of 1:8 with average shoot length of 5.1 cm, following medium gelled with isabgol and sucrose which showed 1:7 multiplication rate with 4.4 cm average shoot length. With the increase in number of passages on medium gelled with isabgol (15g/l), rate of shoot multiplication and shoot length increased till 4th subculture. Maximum number of microrhizome per shoot i.e. 5 was produced on MS medium supplemented with 70 g/l table sugar, followed by 4 microrhizome per shoot on MS medium supplemented with 90 g/l table sugar + 0.9 mg/l BAP, after dark treatment. Rooted plantlets were transplanted on different potting mixture for hardening and kept in the glasshouse. Whereas, maximum per cent survival (90.6%) was observed on cocopeat : sand : FYM (1:1:1). Potting mixture drenched with 20 ml Jeevamrit (3%) and 5 ml of PGPR showed survival percentage of 95 and 41, respectively. Cost analysis of various components showed that there was a significant cost difference between control medium and low cost medium. Control medium was found to be the most expensive resulting in per plantlet cost of Rs. 0.46 whereas, on medium containing isabgol + table sugar and isabgol + sucrose it was Rs. 0.06 and 0.12, respectively. Per plant cost after 4 weeks of primary hardening was calculated to be Rs. 0.71 and Rs. 0.77 for plantlets regenerated on LCM5 and LCM2, whereas it was Rs. 1.11 on control, thus proving that isabgol and table sugar can be the best cheaper alternate to the laboratory grade agar and sucrose.ThesisItem Open Access DEVELOPMENT OF IN VITRO CELL LINES AGAINST Phytophthora nicotianae var. parasitica (DASTUR) WATERHOUSE CAUSING BUCKEYE ROT IN TOMATO (Solanum lycopersicum L.)(UHF,NAUNI, 2020-02) KUMARI, CHANCHAL; SHARMA, RAJNISHABSTRACT Tomato (Solanum lycopersicum L.) crop and yield is suffered every year due to a number of pathogenic diseases. Such diseases are caused by fungi, bacteria, viruses and nematodes which develop through soil-borne, above-ground infections and in some instances are transmitted through insects. The present investigation aimed at development of in vitro cell lines against soil-borne Phytophthora nicotianae var. parasitica (Dastur) Waterhouse causing buckeye rot of tomato. In vitro and in vivo leaf and cotyledon explants of cv. Solan Lalima were used for the callus induction. Highest per cent callus survival and highest average shoot regeneration was obtained in MS medium supplemented with 1.0 mg/l 2,4-D along with 0.5 mg/l Kn using cotyledon as explants and 0.5 mg/l NAA along with 1.0 mg/l BAP using leaf explants, respectively. The developed calli was subjected on selective medium supplemented with 25% culture filtrate of targeted pathogen, where 18.32% average callus survival and 34.59% average shoot induction was recorded. Further confirmation of somaclonal variants was done using RAPD markers. Thus it was concluded that putative somaclonal variants in tomato var. Solan Lalima against buckeye rot pathogen P. nicotianae var. parasitica have been produced and these variants will be further validated by performing various bioassays under field conditions.