1. KAUTIR (Kerala Agricultural University Theses Information and Retrieval)

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    Molecular characterization of Ginger genotypes (Zingiber officinale Rosc.)
    (Department of Plant Biotechnology, College of Agriculture , Vellayani, 2021-12-22) Anaswara, P A; Sreekala, G S
    The study entitled “Molecular characterization of ginger genotypes (Zingiber officinale Rosc.)” was carried out at the Department of Plant Biotechnology and Department of Plantation Crops and Spices, College of Agriculture, Vellayani during 2020 to 2021. The objective of the study was to assess the genetic diversity of ginger genotypes using simple sequence repeats (SSR) and inter-simple sequence repeat (ISSR) markers. Leaf samples of twenty ginger genotypes maintained in the germplasm of Department of Plantation Crops and Spices were collected and subjected for DNA isolation using CTAB method. Quality and quantity of the isolated DNA samples were determined using Agarose gel electrophoresis and spectrophotometric analysis. Fifteen ISSR (ISSR 26, ISSR 14, ISSR 53, ISSR 54, ISSR 69, ISSR 79, ISSR 11, ISSR 12, ISSR 72, ISSR 04, UBC 835, UBC 809, UBC 829, UBC 818 and UBC 828) and 15 SSR (GES 440, GES 452, GES 454, GB-ZOM-033, GB-ZOM-040, GB-ZOM-055, GBZOM-064, GB-ZOM-103, GB-ZOM-107, GB-ZOM-111, GB-ZOM-140, RM 154, RM 171, RM 135, and RM 125) markers were selected from previous studies for molecular characterization of ginger genotypes. Out of these fifteen SSR primers four were rice SSR primers (RM 154, RM 171, RM 135, and RM 125) with high Polymorphic Information Content (PIC) value. Isolated DNA samples showed good quality and their concentration ranged from 132 to 3162 μg/ml. Average polymorphism of ginger genotypes using ISSR and SSR primers were 56% and 67.6% respectively which revealed a moderate level of polymorphism within the twenty ginger genotypes. ISSR markers UBC 829, ISSR 53, ISSR 54, ISSR 72 and SSR markers GB-ZOM-055, GB-ZOM-103, GB-ZOM-064, RM 154, RM 171, RM 135 showed 100 percentage of polymorphic loci. PIC value of ISSR primers ranged from 0 (UBC 818, UBC 828, ISSR 69 and ISSR 12) to 0.49 (UBC 829) and SSR primers ranged from 0 (GES 452, GES 454, GES 440, GB-ZOM111, GB-ZOM-033, GB-ZOM-140) to 0.45 (RM 154). The Mantel’s statistic (r) value based on Spearman's rank correlation obtained as 0.01652 with a significance value (p) of 0.4457 indicated no significant correlation between SSR and ISSR marker information. In this study Jaccard’s similarity coefficients using ISSR and SSR marker data ranged from 0.69 to 0.98 and 0.54 to 0.98. Dendrogram generated using ISSR and SSR data separated twenty genotypes into three clusters and PCoA of twenty ginger genotypes using ISSR and SSR data revealed 11 and 9 clusters respectively. A low to moderate level of polymorphism between these twenty ginger genotypes were observed using ISSR and SSR marker analysis. Divergent lines identified from the Dendrogram of ISSR and SSR data are Mananthavady (T1), Murickassery (T14), Thalavur (T20), Mannarkkad (T9), Plamoodu (T21), Kazhakoottam 1 (T11) and Kazhakoottam 2 (T22). Compared to all other ginger genotypes Plamoodu (T21) and Mannarkkad (T9) showed high variation in ISSR and SSR marker analysis. The marker analysis revealed a higher similarity between ginger genotypes Kottarakkara (T5) and Kothamangalam (T7). The results of the study indicated the efficiency of SSR markers over ISSR markers in evaluating the genetic diversity of ginger genotypes. Polymorphic bands produced by the rice-SSR markers revealed that they are suitable for crossamplification studies in ginger. Among the twenty ginger genotypes characterized using molecular markers, higher variation was noticed in Plamoodu (T21) and Mannarkkad (T9) showing their suitability in future selection programs. The variation detected at genetic level among these ginger genotypes from this study will be useful for future genetic diversity studies.
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    Development of heteroploids in ginger (Zingiber off zingiber officinale Rosc.)
    (Department of Plantation Spices Medicinal and Aromatic Crops, College of Agriculture, Vellayani, 2024-01-24) Silpa, S G; Sreekala, G S
    An experiment entitled “Development of heteroploids in ginger (Zingiber officinale Rosc.)” was carried out in the Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture, Vellayani, during October 2018 to October 2022 with an objective to develop heteroploids using colchicine for yield and quality in ginger cultivars. The present study to develop heteroploids in ginger (Zingiber officinale Rosc.)” was carried out as three experiments (i) Development of heteroploids in ginger cultivars using colchicine in vivo (ii) Development of heteroploids in ginger cultivars using colchicine in vitro and (iii) Screening of heteroploids for stability. Experiments conducted at Department of Plantation, Spices, Medicinal and Aromatic Crops during 2018 recorded four promising genotypes of ginger. Rhizomes of these genotypes along with rhizomes of ginger varieties Athira, Aswathy, IISR Varada and IISR Mahima were collected from College of Agriculture, Vellanikkara and ICAR- Indian Institute of Spices Research respectively and heteroploidy was induced in vivo as per the method developed by Sheeba (1996). A total of sixteen treatments including eight respective controls were tried in Completely Randomized Design.
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    Ginger (Zingiber officinale) yield variability under different climate change scenarios
    (Department of Agricultural Meteorology, College of Agriculture, Vellanikkara, 2021) Fathima Sona, N; Shajeesh Jan, P
    Ginger (Zingiber Officinale) is an important commercial spice crop grown from very ancient times in India. Because of its valued aroma and lemon flavour, it has spread to tropical and subtropical regions from the Indo-China region and became one of the earliest oriental spices known to Europe (Nybe and Miniraj, 2005). Species diversity and yield of spices are threatened due to the ever increasing temperature and changes in precipitation pattern. Since spices are grownboth in plains and high altitudes, it is important to assess the long term climatic changes within a region and its influence on productivity (Sing, 2008). The present experiment was aimed to study the impact of climate change on growth and yield aspects of ginger crop under climate change scenarios of RCP 4.5 and 8.5. Ginger varieties, Maran and Varada were raised at Instructional Farm (IF) Vellanikkara, by adopting split plot design. The experiment was laid out with four dates of planting (15th May, 1st June, 15th June and1 st July) as main plot treatment and two varieties (Maran and Varada) as sub plot treatments. Fourreplications were given for the experiment. The crop weather relationship was analysed with correlation with the help of SPSS software. Principal Component Analysis (PCA), a multivariate statistical technique was done in order to reduce the multicollinarity of large data sets of weather variables to substantially smaller sets of new variables. The developed principal components were utilized for model development using stepwise regression analysis. The future climate was estimated by climate change projections generated using CCSM4 models for 2030, 2050 and 2080based on scenarios RCP 4.5 and 8.5. The life cycle of ginger was characterized by four distinct stages, ie., sowing to 50% germination, 50% germination to active tillering, active tillering to bulking and bulking to physiological maturity. Duration taken for each phenophases found to vary for all the four date of planting in both Maran and Varada. The May 15th date of planting took more days to germinate compared to other date of plantings. The number of days taken for sowing to germination was positively correlated with maximum temperature, minimum temperature, rainfall and soil temperature in both varieties. The number of days taken to attain physiological maturity also foundto be more in May 15th planted crop. The weather experienced during various phenophases have significant influence on yield and other yield attributes of ginger crop. It was found that the yield of both varieties of ginger havepositive correlation with minimum temperature at all the four phenophases except bulking t o physiological maturity. Maximum temperature observed at sowing to germination was positively correlated with yield of both Maran and Varada. At active tillering to bulking stage, rainfall, rainy days and minimum temperature showed a significant positive correlation with yield, but maximum temperature, wind speed and solar radiation showed a significant negative correlation with yield. The Principal Component Analysis was carried out for ginger varieties Maran and Varadaby using the weather parameters experienced in four stages ie., sowing to 50% germination, 50% germination to active tillering, active tillering to bulking and bulking to physiological maturity. Weather parameters considered include maximum temperature (Tmax), minimum temperature (Tmin), rainfall (RF), rainy days (RD), relative humidity (RH), wind speed (WS) and solar radiation (SRAD). The statistical model was developed with principal components as independentand yield as dependent variable. Projected climatic conditions of Vellanikkara, Thrissur under climate change scenarios of RCP 4.5 and 8.5 were downscaled from CCSM4 model. The projected climate of near century (2010-2039), midcentury (2040-2069) and end of century (2070-2099) were downscaled for the study. The projected yield of the ginger variety Maran was found to decrease at all planting datesexcept July 1 st under both the RCP 4.5 and 8.5 scenarios. Under RCP 4.5 scenario, more reductionwas reported at the end of the century on May 15th (38%), June 1st (29%) and June 15th (54.5%) dates of planting. July 1 st planted crop reported increase in projected yield under near, mid and end of centuries. More increase in yield was observed in midcentury (17.7%). Under RCP 8.5 scenario,May 15th (50.2%) and June 1st (20.3%) dates of planting reported the highest percentage of yield reduction during midcentury. June 15th date of planting, recorded the highest yield reduction of 52.6% at the end of the century. July 1st planting date recorded increase in yield and it was more in end of century (15.5%). In case of Varada, under RCP 4.5 scenario, more yield reduction was reported at the end of century during May 15th (24%) and June 1 st (7.2%) dates of planting. DuringJune 15th planting dates, near and end of century reported the same yield reduction of 24.9%. TheJuly 1 st planted crop reported an increase in yield, which was more (15.5%) during end of century. Under RCP 8.5 scenario, more yield reduction was observed in midcentury on May 15th (28.8%) date of planting. June 1st (6.6%) and June 15th (19.9%) dates of planting reported more reduction in end of century. July 1 st reported more increase of 24% in end of century.
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    Evaluation of CO2 enrichment on growth, development and soft rot tolerance in ginger (Zingiber officinale Rosc.)
    (Department of Plant Physiology, College of Agriculture, Vellayani, 2021) Manasa, R; Manju, R V
    The present investigation entitled “Evaluation of CO 2 enrichment on growth, development and soft rot tolerance in ginger (Zingiber officinale Rosc.)” was undertaken with the objective to assess the impact of elevated CO 2 on growth, yield, quality and tolerance to soft rot in ginger through morphological, physiological, biochemical and molecular analysis. Two sets of pot culture experiments were conducted during 2017- 2019. The technology utilized for CO 2 enrichment was Open Top Chamber (OTC) system established under Department of Plant Physiology, College of Agriculture, Vellayani. In the first experiment three varieties of ginger Aswathy, Athira and Maran were utilized to assess the impact of elevated CO 2 on growth, yield and quality of ginger. One set of three month old plants were shifted to OTC and another set was retained under ambient CO 2. The plants were harvested at 4 th , 5 th , 7 th and 8 th months after planting (MAP). Observations on growth, physiological and biochemical parameters were recorded at each harvest. Yield, yield parameters, tissue nutrient status and quality parameters were assessed at 8 MAP. Second experiment was conducted to assess the impact of elevated CO 2 on tolerance to soft rot in three ginger varieties Aswathy, Athira, Maran and wild ginger congener Zingiber zerumbet. One set of three and a half month old plants were shifted to OTC and another set retained at ambient CO 2 and were challenged with Pythium aphanidermatum at 4 MAP. The CO 2 concentration of 500 ppm was maintained in the OTC. The results of the first experiment indicated an improvement in growth parameters like plant height, number of leaves, root weight, shoot weight, stem diameter and number of tillers, physiological and biochemical parameters like total chlorophyll, photosynthetic rate and total sugars in the rhizome by recording significantly higher values upon CO 2 enrichment when compared to ambient CO 2 condition. Significant reduction in transpiration rate, stomatal conductance and specific leaf area were recorded upon CO 2 enrichment. The per cent increase in yield parameters like number of fingers (38.19%), rhizome spread (11.32%), thickness (42.72%) and dry weight of rhizomes (31.43%) recordedhigher values upon CO 2 enrichment, based on average performance of all the selected varieties, leading to greater yield. The per cent increase in the fresh weight of rhizome recorded higher for variety Aswathy (38.34%), followed by Maran (20.00%) and Athira (12.12%) with a total fresh rhizome yield of 165.35 g plant -1 under ambient and 228.75 g plant -1 under elevated CO 2 condition in the case of Aswathy. Upon CO 2 enrichment the quality of rhizome was modified with increase in fiber (60.26%), gingerol (9.06%), phenol (19.25%), total oleoresin (11.52%), starch (44.84%) and essential oils (31.10%). Significant reduction in protein and nutritional status in the rhizomes were also recorded. Nitrogen, potassium, iron and zinc contents decreased by 15.78%, 3.22%, 11.78% and 18.57% respectively. Phosphorous and copper showed decrease in trend though not significant. Among the three varieties of ginger, Aswathy was found to be performing better compared to Athira and Maran not only in terms of yield, also for majority of quality parameters assessed upon exposure to elevated CO 2 condition. In the case of impact of elevated CO 2 on soft rot development, there was no variation in the per cent disease incidence. However, lesser number of days was taken for development of symptom. The per cent disease index was reduced by 11.10% upon CO 2 enrichment. The reduction in the fresh weight of rhizome upon pathogen inoculation was also less under elevated CO 2 (14.34%) when compared to ambient CO 2 (17.60%) showing that increasing CO 2 levels might bring down the disease severity. Upon P. aphanidermatum inoculation, the retention of photosynthetic pigments like chlorophyll a (0.90 mg g -1 FW) chlorophyll b (0.74 mg g -1 FW) total chlorophyll (1.64 mg g -1 FW) carotenoids (0.58 mg g -1 FW), xanthophyll (0.40 mg g -1 FW) and photosynthetic rate (5.92 μmol CO 2 m -2 s -1 ) were higher under elevated CO 2 when compared to ambient CO 2 . Higher membrane integrity values and lower malondialdehyde (0.75 μg g -1 FW) content in all the selected varieties were evident upon pathogen inoculation under CO 2 enrichment. Enhanced activity of ascorbate peroxidase (4.86 activity Units mg -1 total protein), PAL (Phenylalanine ammonia lyase) (20.91 Units mg -1 total protein), and glucanase (4.57Units mg -1 total protein) activity, as well as higher accumulation of phenols (32.40 mg g -1 DW) and ascorbic acid (7.14 mg 100 g -1 FW) were found in the inoculated plants under CO 2 enrichment compared to the ambient CO 2 . The catalase activity was found to reduce upon pathogen inoculation and the reduction was higher (0.11 Units mg -1 total protein) under ambient CO 2 as compared to elevated CO 2 (0.09 Units mg -1 total protein). Highest levels of hydrogen peroxide, superoxide and cell death were observed upon pathogen inoculation under ambient CO 2 through staining techniques and it was highest for variety Aswathy and least for Z. zerumbet. After pathogen inoculation maximum values for variable to maximum fluorescence Fv/Fm (0.63), photosynthetic electron transport rate (82.60 micro equiv m -2 s -1 ) and quantum yield of PS II photochemistry (0.29) were recorded under elevated CO 2. Protein profiling performed in varieties Athira and Aswathy revealed lower expressions of 20-25 kDa and 25-35 kDa protein under pathogen inoculation in variety Aswathy. The expression of 55 kDa (Rubisco Large Sub- Unit) was down regulated to a greater extent under pathogen inoculation in both the varieties. Higher expression levels of the defense responsive genes Phenylalanine ammonium-lyase (PAL), Cinnamyl alcohol dehydrogenase (CAD) and Chalcone synthase (CSH) were also observed upon pathogen inoculation under CO 2 enrichment in both the varieties. Between the varieties, Athira had higher relative fold changes of PAL, CAD and CSH under both ambient and elevated CO 2 upon pathogen inoculation. The present study revealed that CO 2 enrichment can improve the performance of ginger in terms of growth, yield and quality. It also indicates changes in photosynthetic allocation pattern and phytochemical profiles with enhanced contents of oleoresins and essential oils. Ginger plants showed better tolerance against P. aphanidermatum upon CO 2 enrichment. This was apparent in the form of reduced disease severity, oxidative stress level and less reduction in yield upon pathogen infestation under elevated CO 2 condition. This was evidently achieved through the activation of multiple mechanisms, including enhanced production of anti-oxidants and up regulation of pathogen resistance proteins. This is one of the first studies to comprehend on possible effects of elevated CO 2 on the susceptibility of ginger to soft rot. The study also demonstrated the varietal variations existing in the response to CO 2 enrichment which will help in selection ofsuitable varieties for changing climatic scenario. The information generated indicates the possibility of enhancing both qualitative and quantitative yield of ginger through low cost CO 2 enrichment technologies. The insights obtained on the tolerance mechanisms involved will help to restructure the soft rot disease management strategies in the coming future.
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    Commercial production of ginger (Zingiber officinale ROSC) microrhizomes using temporary immersion bioreactor system
    (Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2020) Rasha Fathima, A A; Shylaja, M R
    Ginger (Zingiber officinale Rosc.), is an important commercial spice crop grown in India from very ancient times. High seed rate of ginger (1500 kg/ha) and desiccation of seed rhizomes during storage are the problems faced by farmers in ginger cultivation. The tissue culture plants are not commercially distributed in ginger as they require an additional one more season for rhizome formation. Microrhizomes of ginger induced in vitro if used as planting materials, rhizomes can be harvested in the same season as conventional seed rhizomes and year round availability of seed material can be ensured. Hence, in vitro induced microrhizomes are included in the seed chain of ginger and there is high demand for microrhizomes for clean ginger production. The study entitled “Commercial production of ginger (Zingiber officinale Rosc.) microrhizomes using Temporary Immersion Bioreactor (TIB) system” was conducted at Centre for Plant Biotechnology and Molecular Biology (CPBMB), College of Horticulture, Vellanikkara during 2018 to 2020. The objective of the study was to develop an efficient commercial production protocol for ginger microrhizomes using a TIB system. The study was conducted in the ginger variety Athira using Plantform TIB purchased from Sweden. The multiple shoot cultures in the 5th subculture stage received from the commercial micropropagation unit, of CPBMB were used for the study. The protocol for in vitro induction of microrhizomes reported by Shylaja et al. (2016) was optimised for bioreactor production and at each stage it was compared with the conventional microrhizome production. The number of clumps/ 500ml of medium to initiate multiple shoot production, media for shoot multiplication and microrhizome induction were optimised for TIB and compared with conventional microrhizome production system. The microrhizome, root and shoot characters in microrhizome plantlets and growth of microrhizome plants after hardening were evaluated in the two culture systems. The clonal fidelity analyses of microrhizome plants derived from 8th subculture cycle were done using the specific ISSR marker as reported by Gavande, (2013). The clump size of 15 clumps/ 500 ml of medium showed higher shoot multiplication in both TIB and conventional system. The shoot proliferation in bioreactor (7.71 shoots/clump) was significantly higher than the conventional microrhizome production system (5.24 shoots/clump). Early induction of microrhizomes was observed in TIB system. In both the culture systems, microrhizome induction was faster in MS medium with 90 gL-1 sucrose. The number of microrhizomes produced in the medium in TIB varied from 87.75 to 96.75/ 500ml medium and in conventional system it varied from 84 to 88/ 500 ml medium. The microrhizome plantlets produced in TIB recorded significantly higher weight of microrhizomes, better shoot and root growth and more number of leaves compared to the microrhizome plantlets produced in the conventional system. The weight of microrhizomes, number of roots and root length were higher in microrhizome plantlets in MS medium with 90 gL-1 sucrose. The mean weight of microrhizome in the medium was 0.29 g in bioreactor and 0.18 g in conventional system. The survival of microrhizome plants after primary hardening was 94.34 per cent and after secondary hardening was 99.29 per cent in TIB. Microrhizome plants from bioreactor recorded significantly higher plant height (18.60 cm) compared to conventional system (14.80 cm). The number of leaves recorded in microrhizome plants of both the culture systems were on par and ranged from 6.97 to 7.18. Clonal fidelity analyses using specific ISSR marker revealed that there were no polymorphism in the ISSR amplification profiles in microrhizome plants produced after the 8th subculture cycle with the source mother plant and hence plants produced from both the culture systems are true to type. The protocol developed in the present study can be further modified by early bulking of the clumps, employing TIB for multiple shoot production in early culture phase and reducing the number of culture cycles so that the entire protocol period can be reduced. Evaluation of TIB microrhizome plants in high tech poly house or field and comparison of yield and quality with microrhizome plants from conventional production system also can be focused for further research.
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    In vitro synthesis of Gingerol and analysis of expressed sequence tags for Gingerol production in Ginger(Zingiber officinale Rosc.)
    (Centre for plant biotechnology and molecular biology,College of Horticulture, Vellanikkara, 2020) Manjusha, Rani; Shylaja, M R
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    Study on the effect of foliar application of urea and planofix on the growth, yield and quality of ginger varieties (Zingiber officinale Roscoe)
    (Department of Horticulture, College of Agriculture, Bhubaneswar, 1975) Sreekandan Nair, G; Das, R C
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    Induction of autotetraploid in ginger
    (Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 1996) Sheeba, P T; Alice Kurian
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    Ginger direct procurement programme of synthite industries limited: A study on direct sourcing
    (College of co-operation, banking and management, Vellanikkara, 2016) Amalraj, K R; Binoo P Bonny
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    Yield improvement in tranplanted ginger by seed priming and biostimulant spray
    (Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2020) Ann Sneha Baby; Jalaja S Menon