Thumbnail Image

Kerala Agricultural University, Thrissur

The history of agricultural education in Kerala can be traced back to the year 1896 when a scheme was evolved in the erstwhile Travancore State to train a few young men in scientific agriculture at the Demonstration Farm, Karamana, Thiruvananthapuram, presently, the Cropping Systems Research Centre under Kerala Agricultural University. Agriculture was introduced as an optional subject in the middle school classes in the State in 1922 when an Agricultural Middle School was started at Aluva, Ernakulam District. The popularity and usefulness of this school led to the starting of similar institutions at Kottarakkara and Konni in 1928 and 1931 respectively. Agriculture was later introduced as an optional subject for Intermediate Course in 1953. In 1955, the erstwhile Government of Travancore-Cochin started the Agricultural College and Research Institute at Vellayani, Thiruvananthapuram and the College of Veterinary and Animal Sciences at Mannuthy, Thrissur for imparting higher education in agricultural and veterinary sciences, respectively. These institutions were brought under the direct administrative control of the Department of Agriculture and the Department of Animal Husbandry, respectively. With the formation of Kerala State in 1956, these two colleges were affiliated to the University of Kerala. The post-graduate programmes leading to M.Sc. (Ag), M.V.Sc. and Ph.D. degrees were started in 1961, 1962 and 1965 respectively. On the recommendation of the Second National Education Commission (1964-66) headed by Dr. D.S. Kothari, the then Chairman of the University Grants Commission, one Agricultural University in each State was established. The State Agricultural Universities (SAUs) were established in India as an integral part of the National Agricultural Research System to give the much needed impetus to Agriculture Education and Research in the Country. As a result the Kerala Agricultural University (KAU) was established on 24th February 1971 by virtue of the Act 33 of 1971 and started functioning on 1st February 1972. The Kerala Agricultural University is the 15th in the series of the SAUs. In accordance with the provisions of KAU Act of 1971, the Agricultural College and Research Institute at Vellayani, and the College of Veterinary and Animal Sciences, Mannuthy, were brought under the Kerala Agricultural University. In addition, twenty one agricultural and animal husbandry research stations were also transferred to the KAU for taking up research and extension programmes on various crops, animals, birds, etc. During 2011, Kerala Agricultural University was trifurcated into Kerala Veterinary and Animal Sciences University (KVASU), Kerala University of Fisheries and Ocean Studies (KUFOS) and Kerala Agricultural University (KAU). Now the University has seven colleges (four Agriculture, one Agricultural Engineering, one Forestry, one Co-operation Banking & Management), six RARSs, seven KVKs, 15 Research Stations and 16 Research and Extension Units under the faculties of Agriculture, Agricultural Engineering and Forestry. In addition, one Academy on Climate Change Adaptation and one Institute of Agricultural Technology offering M.Sc. (Integrated) Climate Change Adaptation and Diploma in Agricultural Sciences respectively are also functioning in Kerala Agricultural University.

Browse

2013 - 20174
Reset filters
Subject: Plant Biotechnology × Author: KAU × End date: 2017 × Type: Thesis × Thesis Type: Ph.D ×
Reset filters

Search Results

Now showing 1 - 4 of 4
  • Thumbnail Image
    ThesisItemOpen Access
    Metabolite Profiling and gene expression analysis for gingerol production in selected somaclones of ginger (zingiber officinale rosc.)
    (Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2017) Sreeja, S; KAU; Shylaja, M R
    Ginger (Zingiber officinale Rosc.) is one of the oldest known spices and is much valued for its medicinal properties. Ginger rhizome is a source of active biological compounds which are responsible for its medicinal properties. Gingerol is the major pungent polyphenol in ginger which has got a wide array of pharmacological properties. Research on somaclonal variation done at Kerala Agricultural University could locate ginger somaclones with high gingerol content and in the course of investigations high somaclonal variation was observed for quality components. Ginger has a very big genome of 23,618 Mbp which is little exploited and reports on the genomic base of gingerol biosynthesis are scanty. The present investigations hence aim at profiling the metabolites in selected ginger somaclones using high throughput analytical platforms and to analyze the gene expression with respect to gingerol production. One released ginger variety from KAU (Athira), two selected ginger somaclones (B3 and 132M) and parent cultivar (Maran) formed the experimental materials for the study. Studies were carried out at Centre for Plant Biotechnology and Molecular Biology, Distributed Information Centre of College of Horticulture and Arjuna Natural Extracts Pvt. Ltd., Aluva during August 2013 to July 2017. The profiling of aroma principles using Gas Chromatography-Mass Spectrometry (GC-MS) and pungency principles using High Performance Liquid Chromatography (HPLC) at various growth stages viz. five months after planting (5MAP), six months after planting (6MAP) and seven months after planting (7MAP), revealed that aroma and pungency principles accumulated in ginger rhizomes at the rhizome formation stage (5MAP). Clone to clone variation was observed in the number and quantity of aroma and pungency principles accumulated in the rhizome. Total gingerol content in somaclone B3 (19.07%) was high when compared to the control cultivar Maran (17.49%) irrespective of the growth stages. Gene expression for Chalcone synthase in selected somaclones done using real time PCR assay showed highest gene expression in somaclone B3 when the control cultivar Maran was set as calibrator. Somaclone B3 recorded 54 per cent increase in Chalcone synthase gene expression over the control cultivar Maran. Suppression subtractive hybridization done to identify differentially expressed genes in somaclone B3 and control cultivar Maran could prepare Expressed Sequence Tag (EST) libraries both for rhizome and leaf. Analysis of EST sequences (25 rhizome ESTs and 19 leaf ESTs) using various bioinformatic tools revealed that there were no differentially expressed genes for gingerol production in rhizome ESTs. But eleven differentially expressed proteins involved in signaling response, protein trafficking, photosynthesis, ATP formation and transposon mediated mutation were observed in rhizome ESTs. The analysis of leaf ESTs showed differential gene expression in somaclone B3 for 3-ketoacyl CoA thiolase (ACAA1) gene which is involved in gingerol biosynthetic pathway. Hence the higher expression of 3-ketoacyl CoA thiolase gene is responsible for the high gingerol content in somaclone B3 as compared to control cultivar Maran. Eighteen other differentially expressed proteins involved in biological processes like transportation of plant secondary metabolites and their intermediates, mobilization of sucrose into pathways involved in metabolism, lipid biosynthesis, transportation of cellular material to microtubules, biogenesis of metabolic pathways in Calvin cycle were observed in leaf ESTs. The differentially expressed gene (ACAA1) can be further validated using northern blotting and quantitative real time PCR by designing specific primers from the ESTs. Expressed sequence tags and corresponding differentially expressed proteins can be used as molecular markers. Post translational modification in differentially expressed proteins can be used to study the mechanism of gingerol production. Forty four sequences deposited at NCBI form the base sequences for further research.
  • Thumbnail Image
    ThesisItemOpen Access
    Development of small interfering RNA (siRNA) mediated resistance in banana against banana bract mosaic virus
    (Department of Plant Biotechnology, College of Agriculture, Vellayani, 2016) Lekshmi, R S; KAU; Soni, K B
    The present study entitled “Development of small interfering RNA (siRNA) mediated resistance in banana against Banana bract mosaic virus (BBrMV)” was carried out during 2012-2016 in the Department of Plant Biotechnology, College of Agriculture, Vellayani. The study was carried out with an objective to develop siRNA mediated technology for the development of banana plants resistant to Banana Bract Mosaic Virus (BBrMV). The study was conducted in banana cv. Nendran. A protocol for somatic embryogenesis in banana cv. Nendran was standardized by using immature male flowers as explants. Pale white friable callus with rich cytoplasm was initiated in Murashige and Skooge (MS) medium supplemented with BA (0.1 – 0.5 mgL-1) and picloram (0.5 – 2 mgL-1) incubated in dark with a maximum explant response of 30 per cent. For embryogenesis, the developed embryogenic calli were transferred to semisolid MS medium supplemented with BA 2 mgL-1 and IAA 0.5 mgL-1 which resulted in a maximum of 10 per cent embryogenesis. The glassy elongated monocot embryos were germinated in half strength semisolid MS medium (0.3 per cent Gelrite) supplemented with BA 2 mg L-1 and IAA 0.5 mg L-1 and incubated in dark. A maximum germination rate of 80 per cent was obtained in this medium. The germinated embryos were transferred to MS medium with BA 2 mg L-1 and NAA 1 mg L-1 resulted in 100 per cent Plant regeneration. The plantlets were transferred to coirpith compost in pot trays in mist chamber for one month for hardening and then transferred to polybags with soil and cowdung (1:1) mixture. To develop siRNA technology to silence the replicase gene of BBrMV, an intron hairpin RNA (ihpRNA) construct was developed. For this a partial mRNA sequence of replicase gene was isolated from BBrMV banana plants. Gene specific primers designed based on the whole genome sequence information retrieved from the GenBank, NCBI. Total RNA from infected banana leaves was isolated and cDNA was prepared using RT-PCR. The partial gene fragment isolated was sequenced and analysed using the bioinformatics tool BLAST. The sequence was subjected to miRNA target prediction and restriction mapping to select suitable region for the construct and further processing. Based on this information a fragment of 400 bp towards the 5’ end was amplified by designing a set of primers with anchored restriction sites. The primers anchored with BamHI and PacI sites were used for the amplification of sense strand and primers anchored with KpnI and SpeI sites were used for antisense strand amplification. The sense and antisense fragments amplified were cloned to pTZ57R/T cloning vector. In the next step the inserts were released from pTZ57R/T using the corresponding restriction enzymes and were integrated in pSTARLING (primary vector), on either side of the cre intron which facilitated the formation of the hairpin (ihpRNA) construct. Presence of the inserts was confirmed by restriction digestion and electrophoresis. The ihpRNA construct in pSTARLING now contained ubiquitin promoter, ubiquitin intron, sense strand of replicase gene, cre intron, antisense strand of replicase and termination sequence in the order with the NotI restriction sites. This construct was released from pSTARLING and ligated to the digested NotI site in the lacZ gene of the binary vector pART27 containing antibiotic resistance marker nptII and spec. The binary vector was confirmed for the insert by transferring to DH5α and colony selection by blue-white screening. The binary vector with the insert isolated from the white colony, was transferred to Agrobacterium tumefaciens strain LBA 4404 via freeze-thaw method. Transformed colonies were picked up and confirmed the presence of the vector and the ihpRNA insert by PCR. Somatic embryos were transformed with LBA 4404 carrying the ihpRNA construct the ihpRNA construct and the transformed embryos were selected with antibiotic pressure (Kanamycin 100 mg L-1). Transformed embryos were subjected to regeneration. A maximum regeneration of 25 per cent was obtained after transformation. The regenerants were confirmed for the presence of ihpRNA construct using PCR with specific primers for sense-intron-antisense fragment, npt II and cre intron. The study was successful in developing a siRNA construct for resistance against BBrMV and obtaining transformed Nendran banana plantlets.
  • Thumbnail Image
    ThesisItemOpen Access
    Validation of apomixis and transcriptome analysis for detection of the genes related to apomixis in black pepper (piper nigrum L.)
    (Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2017) Rohini Rajkumar, Bansode; KAU; Valsala, P A
    Black pepper (Piper nigrum L.) universally honoured as “Black Gold” and also known as “King of Spices” is one of the most important spices in the world. It is a perennial climber belonging to the family Piperaceae and is valued throughout the world for its spice value and medicinal properties. Black pepper is grown in Southern India mainly in Kerala. Here productivity is declining due to various reasons and continuous vegetative propagation leads to accumulation of diseases and finally results in unproductive vines. Therefore, use of quality planting material of improved varieties is necessitated for the enhancement of productivity. Apomixis is a mode of asexual reproduction where the sexual organs are utilized, but the seeds develop without fertilization. It combines the advantage of seed propagation and vegetative propagation, and can be utilized for developing disease free planting material without losing the clonal integrity. The present study on “Validation of apomixis and transcriptome analysis for detection of the genes related to apomixis in black pepper (Piper nigrum L.)” was undertaken at the Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara during the period 2011-2014 with the objective to validate apomixis in black pepper varieties, Panniyur-1 and Panniyur-2 through controlled pollination studies and to identify differentially expressed genes associated with apomixis through transcriptome analysis. Studies were conducted on bush pepper plants of the selected varieties maintained in the green house. Floral biology of Panniyur-1 and Panniyur-2 were studied. The female phase in Panniyur-1 started 8th day after spike initiation and completed on 27th day. Stigma became receptive on 9th day and receptivity was indicated by creamy white colour. Beginning of male phase indicated by anther emergence on either side of the ovary occurred on 19th day of spike emergence and active dehiscence of anthers occurred on 20th day. In Panniyur-2, beginning of female phase started on 14th day after spike initiation and stigma became receptive on 15th day. Beginning of male phase occurred on 22nd day of spike emergence and active dehiscence of anthers occurred on 23rd day. So in Panniyur-1 and Panniyur-2 the active female and male phase is separated by 10 and 8 days, respectively. Attempts were made to confirm apomixis in Panniyur-1 and Panniyur-2 varieties by allowing berry development in controlled condition by bagging of spike initials. Berry development occurred under bagged condition. In this study, upper six berries were considered as apomictic and lower six berries were considered as pollinated. Efforts were made to regenerate apomictic and non- apomictic progenies under in vitro and ex vitro. In vitro embryo culture resulted in embryo germination and multiple shoot induction in SH medium with hormones. But plants were lost due to microbial contamination and phenolic interference. So ex vitro germinated seedlings were used for further studies. Histological examination of pollinated and unpollinated ovaries was done through microtomy. The results revealed that in case of pollinated berries, the sexual fertilization occurs and embryo develops inside the embryo sac in the micropylar end. Whereas, in apomictic embryo sac more than one aposporous initial cells were observed which were arising from a somatic cell located in the nucellus. So, it can be concluded that facultative apomixis exists in black pepper varieties P1 and P2 and the embryo develops parthenogenetically. The molecular characterization of apomictic and non-apomictic seedlings was done through Inter Simple Sequence Repeats (ISSR) and Simple Sequence Repeats (SSR) assay. Isolation of good quality genomic DNA from apomictic, pollinated progenies and mother plant was carried out from the young leaves using modified Rogers and Bendich (1994) as reported by Mogalayi (2011). In ISSR and SSR assay certain primers showed polymorphism among mother plant, apomictic and pollinated progenies. Similarity matrix was calculated and corresponding dendrogram was also constructed for both markers using UPGMA cluster analysis for P1 and P2 varieties. In ISSR assay the similarity coefficient between P1 mother plant and apomictic progenies ranged from 69 to 89 per cent while it was 64 to 66 per cent between mother plant and pollinated progenies. In P2 variety similarity coefficient between mother plant and apomictic progenies ranged from 76 to 90 per cent while it was 62 to 69 per cent between mother plant and pollinated progenies. In SSR assay similarity coefficient between P1 mother plant and apomictic progenies ranged from 50 to 100 per cent while it was 71 to 100 per cent between pollinated progenies and mother plant. Similar results were obtained for P2 variety also. So apomictic progenies may differ from mother plant in certain characters. RNA mediated transcriptome analysis of the apomictic and pollinated berries was done to detect differentially expressed genes. Good quality RNA was isolated by modified LiCl precipitation method. Total RNA from the apomictic and pollinated berries were taken for DDRT-PCR analysis. The first strand cDNA was synthesized from the above RNA samples using HT11C (AAGCTTTTTTTTTTTC). Each first strand cDNA was used for the second strand amplification with 8 different arbitrary primers. The PCR product was resolved in 6 per cent denaturing urea polyacrylamide gel and visualized after silver staining. The differentially expressed cDNA fragments were retrieved from the gel and reamplified and electrophoresed. The agarose gel electrophoresis showed 7 transcript derived fragments (TDFs) ranging from 200-600 bp. TDFs were cloned using pGEMT vector and were sequenced by outsourcing. The sequence data were analysed by BLASTn and BLASTx. The sequences showed homology to NADH dehydrogenase subunit J, acetyl-CoA-benzylalcohol acetyltransferase and purine permease 4 but no significant sequence similarities for apomictic genes deposited in NCBI database were found.
  • Thumbnail Image
    ThesisItemOpen Access
    Gene expression analysis in relation to Fusarium wilt resistance in banana (Musa spp.)
    (Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2013) Jusna Mariya, P L; KAU; Keshavachandran, R
    Banana is one of the important fruit crops of India. Banana is susceptible to several fungal pathogens, nematodes, viruses and insect pests. The greatest threats to global banana production is Fusarium wilt or Panama wilt caused by Fusarium oxysporum f. sp. cubense. Control of the pathogen is difficult and mainly involves the use of disease free suckers. Although disease resistance exists in some banana cultivars, introducing resistance into commercial cultivars by conventional breeding is difficult due to its triploid nature and sterility factors of banana. The study entitled "Gene expression analysis in relation to Fusarium wilt resistance in banana (Musa spp.)" was carried out at the Centre for Plant Biotechnology and Molecular Biology, Vellanikkara during the period 2009-2013 with an objective to identify differentially expressed genes in disease resistant genotype of banana, Palayankodan using the molecular technique called suppression subtractive hybridization (SSH). Total RNA and mRNA were isolated from healthy and inoculated plants (with Fusarium oxysporum f.sp. cubense) and were used respectively as 'driver' and 'tester' in SSH reaction. The reactions were performed utilizing the PCR select" cDNA subtraction kit provided by CLONTECH, USA. Control subtraction was carried out first using PCR select" cDNA subtraction kit, which gave satisfactory and expected results. For experimental subtraction, the double stranded cDNAs synthesized from Zug mRNA from normal 'driver' and treated 'tester' were digested with RsaI enzyme. Two tester populations were created and each ligated to two different adaptors. This was followed by two hybridization reactions and finally a selective PCR amplification. Only differentially expressed cDNAs were amplified exponentially. This was confirmed by analyzing the PCR products on agarose gel, which showed a smear ranging from 0.9 to 1.3 kb in the subtracted sample and was different from smear pattern of unsubtracted ones. The cDNA fragments from subtracted sample were cloned in pJET and pGEMT vectors and sequenced. Fifty clones were sequenced and analysed after vector and adaptor editing. In silica analysis using bioinformatics tools revealed that some of the cloned sequences showed similarity with known sequences which play important roles during disease resistance conditions directly or indirectly. These included resistance gene candidate NBS type protein, mitogen activated protein kinase, phytoene desaturase, glycerol 3-phosphate dehydrogenase, neutral invertase, 1- aminocyclopropane-l-carboxylase synthase, superoxide dismutase, MADS-box protein, ubiquitin 2, actin, NADPH oxidase, phytoene synthase, ACC synthase, sucrose phosphate synthase, phosphatidic acid phosphatase-like protein, ORF III like polyprotein, bHLH transcription factor like protein, cytochrome oxidase, isochorismatase hydrolase, basic helix-loop-helix family protein, constitutive triple response I-like protein, granule bound starch synthase, alpha amylase precursor, rop protein, GTPase family protein, S-adenosyl-L-methionine synthase protein, ADP-glucose pyrophosphorylase glucose-l-phosphate adenylyl trans, ethylene signal transduction factor and ribosomal protein. Clones were classified into 6 major groups based on function of protein. Sequences had conserved domains for the above mentioned proteins. Genes involved in defense, signal transduction, metabolism, hypothetical protein, transcription factor and translation. For further exploitation of these sequences it is necessary to clone full length cDNA. ESTs thus generated in the present study will be of great use in future for further downstream applications.