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ThesisItem Open Access Molecular characterization of piper species usiing RAPD technique(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2002) Murugan, C; KAU; Sujatha, V SThe study on "Molecular characterization of Piper species using RAPD techniques" was conducted in the Department of Plantation Crops and Spices and CPBMB, College of Horticulture, Vellanikkara, Thrissur from December, 1999 to August, 2001. Material included 13 species/accessions of the genus Piper including Piper nigrutn Linn. These were analysed with 20 selected oligomer decamer primers using PCR technique and agarose gel electrophoresis. The selection of ideal DNA isolation technique was done based on observations of purity of bands, quality of the DNA bands and quantity of DNA recovered. The protocol of Edwards et al. (1991) was selected for DNA isolation with additional step of chloroform-isoamyl alcohol treatment. Fifty decamer primers were screened using DNA. of P. nigrum and 20 were selected based on the number of bands produced, reproducibility of the bands and distinctness of the RAPD profile. RAPD profile of the 13 species of Piper were compared for genetic similarity index (using Jaccard's coefficient), with all 20 selected primers separately. Pooled similarity of the twenty primers put together was found out for 9 species which gave amplification for all the 20 selected primers. The results of pooled analysis was subjected to cluster analysis (SAl-IN) and phenetic dcndrogram was constructed employing UPGMA. Four clusters were obtained, comprising of two accessions of P. nigrum In the first, two accessions of P. longum in the second, P. colubrinutn and P. attenuatum forming the third cluster and P. chaba, P. belle and P. arboreum forming the fourth.ThesisItem Open Access Evaluation of curry leaf (Murraya koenigii L.) accessions for yield and quality(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2019) Anila, Peter; KAU; Krishnakumary, KCurry leaf (Murraya koenigii L.) is an important perennial herbal spice widely used for culinary purpose in India for centuries and the leaves valued for its characteristics flavour and aroma. Leaves possess medicinal and neutraceutical properties due to the presence of vitamins, minerals and phytochemicals. In Kerala, cultivation is limited to landraces which are often reported to be low in quality. Identification of promising types with high yield and quality will pave way for promoting large scale cultivation in Kerala. The present study entitled “Evaluation of curry leaf (Murraya koenigii L.) accessions for yield and quality” was undertaken with objectives of morphological and biochemical characterization of available accessions and standardization of vegetative propagation. accessions of curry leaf maintained by the Department of Vegetable Science, College of Horticulture, Vellanikkara along with the released variety Suvasini were used as the experimental material. Among the accessions, wide variability was recorded for fifteen quantitative and five qualitative morphological characters studied. Cluster analysis was performed and superior accessions were selected and those were subjected to biochemical analysis. Among the accessions, 76 % were having green midrib and petiole whereas 24 % with light red pink. Leaf colour was found as dark green in 19 % of accessions and the remaining accessions were green in colour. Only two leaflet shape were noticed with 19 % having lanceolate shape. The plant height varied from 2.23 m (MK 107) to 5.21m (MK 103) whereas branching height ranged from 31.25 cm (MK 125) to 67.51 cm (MK 122). Accession MK 118 recorded highest canopy spread in N-S direction (4.57 m) while MK 142 recorded in E-W direction (4.78 m). Considerable variability was observed in number of primary, secondary and tertiary branches and more than 40 numbers of tertiary branches were recorded in five accessions namely MK 107, MK 118, MK 126, MK 142 and MK 160. The leaf length ranged from 14.71 cm (MK 122) to 21.81 cm (MK 126) whereas maximum number of leaflets (22 leaflets) were observed in three accessions viz., MK 126, MK 142 and Suvasini with the largest leaf area of 66.46 cm2 in MK 142. Curry leaf was harvested at four months interval from June to March. Harvesting during June- July months in the rainy season recorded the highest leaf yield per plant. Leaf yield of more than 5 kg per plant was obtained in four accessions namely, MK 107, MK 126, MK 142 and MK 160. Aroma and flavour are the important parameters in curry leaf and based on sensory evaluation, eight accessions were identified as superior with a mean sensory score of above 7, with the highest score of 7.40 and 7.33 for flavour and aroma in MK 126. Correlation analysis revealed significant and positive correlation of yield with number of primary, secondary and tertiary branches, canopy spread in N-S and E-W direction, number of leaflets, and leaf area. Based on cluster analysis, all the thirty accessions under the study were grouped into twelve clusters and ten accessions from superior clusters were selected for biochemical analysis. Oil recovery was higher in dry leaves than in fresh whereas antioxidant activity and beta carotene content were higher in fresh leaves. Highest essential oil recovery was recorded in dried samples of accession MK 186 (1.09 %) and maximum content of crude fibre was recorded in two accessions namely MK 118 (11.61 %) and MK 186 (11.65 %). Accession MK 126 recorded the highest content of carbohydrate (15.99 %) and protein (6.00 %). Highest content of total phenol (0.31 %) and antioxidant capacity (3.45) was recorded in MK 105. The beta carotene content was found higher (˃ 8.80 mg 100g-1) in three accessions namely, MK 105, MK 142 and MK 186. Considering yield and quality MK 142 and MK 126 were identified as most promising types followed by MK 105 and MK 186. Standardization of vegetative propagation was done in Suvasini with softwood, semihardwood and hardwood types of cuttings, two growth regulators (IBA and NAA) at different concentrations of 1000ppm, 2000ppm and their combinations during rainy, winter and summer season. Even though success rate of vegetative propagation through stem cuttings was very low, hardwood cuttings treated with highest concentration of growth regulator (2000ppm IBA+2000ppm NAA) resulted in 3.3 % of establishment during rainy seasonThesisItem Open Access Collection and evaluation of Wayanad turmeric (curcuma longa l.) for yield and quality(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2019) Anu, T S; KAU; Jalaja Menon, STurmeric (Curcuma longa L.) is a rhizomatous perennial herb and an important sacred spice of India. It is used as a spice, colourant and an aromatic stimulant in several medicinal preparations. Wayanad local turmeric is reported to contain an average of 9.4 per cent curcumin. Due to the advent of improved varieties, the traditional cultivars of Wayanad are eroding. Keeping in view of this fact the study was undertaken to evaluate the local Wayanad accessions for yield and quality. Twenty six accessions of traditional ecotypes were collected from three taluks of Manathavady, Vythiri and Sultanbathery of Wayanad district through mass publicity. Seventeen accessions having sufficient planting materials for replicated trial were selected for field trial along with three improved varieties, Sona, Kanthi and Prathibha as checks. The crop was raised under the partial shade of 54.29 per cent in the coconut farm of the Department of Plantation crops and spices in randomized block design. Morphological observations were taken at 150 days after planting. Wayanad local turmeric accessions performed better than the check in all the biometric characters except length of the leaf. The plant height was highest for WCL 23 and girth was highest for WCL 3. The number of leaves on the main tiller was significantly high in WCL 15 and 16 while number of tillers per clump was highest in WCL 22. Leaf with maximum length and width were recorded in Kanthi and WCL 16 respectively. Horizontal rhizome branching was observed in the Prathibha , 3, 13, 16, 17, 24 and 25. The variety Prathibha and accessions 16, 17, 20, 21, 24 and 25 showed plumpy rhizomes. Compact rhizomes were observed in 8, 9, 13, 14,15, 16, 19, 20 and 25. The inner core colour of the rhizome was light yellow in accession WCL 24 and reddish yellow Colour was observed in Prathibha, Kanthi and WCL 3. All other accessions had orange inner core colour. The accessions and varieties were categorized based on percent disease index. The variety Prathibha and accessions 3, 13, 14, 15, 16 and 25 were highly resistant to leaf blotch. Accession WCL 5 was highly susceptible to leaf blotch. All the accessions were resistant in disease reaction to leaf spot except WCL 19, which was moderately resistant. The incidence of pests like lema beetle and shoot borer were observed in all accessions while leaf roller incidence was noticed only in Prathibha, WCL 5, WCL 17 and WCL 24. Fresh rhizome yield per plant was significantly high in accessions WCL 24 (350.33 g) and WCL 23 (350.08 g) and were on par with Kanthi (335.60 g) and WCL 25 (274.66 g). Accession WCL 25 was superior in fresh rhizome yield per plot and per hectare (32.32 t ha-1). Accessions WCL 23 and WCL 25 performed over and above the checks in terms of fresh yield, curcumin content and curcumin yield. Curcumin content was high for WCL 5 (10.18 %) followed by WCL 25 (9.27 %) and WCL 26 (9.31 %). The accession WCL 5 was highly susceptible to leaf blotch and curcumin yield (286.06 kg ha-1) was low. The fresh rhizome yield was highest in WCL 25 (32.32 t ha-1). Curcumin yield was highest for WCL 25 (551.40 kg ha-1) and accessions WCL 20, WCL 23, and WCL 26 were on par with WCL 25. Oleoresin content was significantly high in accessions WCL 3 (19.90 %) and WCL 22 (19.57 %). Essential oil content was highest for Sona (4.00 %) and Prathibha (4.00 %). Selection index was also calculated from fresh rhizome yield per clump curcumin content, dry rhizome recovery and disease reaction. Accessions 3, 13, 14, 15, 16, 23 and 25 had a highest score of 12 and these were identified as superior accessions which can be used for further crop improvement programme.ThesisItem Open Access Nutritional and Antioxidant potential of medicinal rice variety Rakthashali(Department of Community Science, College of Horticulture,Vellanikkara, 2019) Aiswarya, Girijan; Aneena, R RThe study entitled “Characterization of viruses infecting small cardamom (Elettaria cardamomum Maton) and production of disease free plants” was conducted at Department of Plant Pathology, College of Agriculture, Vellayani during 2017-2019, with the objective to study the occurrence and distribution of viruses infecting small cardamom, molecular characterization of the viruses and elimination of viruses through meristem culture for the production of disease free planting material. Survey was conducted in Kattappana, Pampadumpara and Ambalapara panchayats of Kattappana block; and Nedumkandam, Thukkupalam and Chembalam panchayats of Nedumkandam blocks of Idukki district during November 2018 – May 2019. The incidence of katte disease caused by Cardamom mosaic virus (CdMV) was present in all the panchayats surveyed and it ranged from 3.75 to 43.0 per cent in Kattappana block and 5.0 to 31.33 per cent in Nedumkandam block. Disease incidence for chlorotic streak caused by Banana bract mosaic virus (BBrMV) was recorded from Kattappana (41%), Pampadumpara (30%) and Nedumkandam (8.33%) panchayats. The aphids infestation was absent in all the surveyed plots. Colocasia spp. and Alpinia spp. were the major plants observed in and around the cardamom fields and were not having visible symptoms of the viral infections. The virus inoculums were maintained under insect proof net house at Cardamom Research Station, Pampadumpara. Katte disease produced slender chlorotic flecks developing into pale green discontinuous stripes running parallel to veins from midrib to leaf margin of the infected leaves. Mosaic mottling and chlorotic specks were seen on the infected leaves and young pseudostems. In case of severe infection, plants produced stunted tillers. Chlorotic streak disease was characterised by continuous and discontinuous chlorotic streaks along veins and midribs of the infected leaves and green discontinuous spindle streaks on pseudostem. CdMV (a potyvirus) and BBrMV in cardamom was detected using polyclonal antibodies of Potato virus Y (PVY) and BBrMV respectively procured from DSMZ, Germany by direct antigen coating- Enzyme linked immunosorbent assay (DAC-ELISA) and Dot immunobinding assay (DIBA).The highest virus titre of CdMV and BBrMV was obtained in samples collected from Pampadumpara and Kattappana respectively. Molecular detection of the viruses was carried out using reverse transcriptase - polymerase chain reaction (RT-PCR) with specific primers for CdMV and BBrMV; and obtained amplicons of expected size of 879-905 bp for CdMV- and 625-633 bp for BBrMV- infected samples. The sequences of the isolates of CdMV from Kattappana, Pampadumpara and Nedumkandam were subjected to BLAST analysis and found to be similar to Indian cardamom mosaic virus isolates from Thalathamane and Appangala with > 96 per cent similarity. The BBrMV in cardamom from Kattappana, Pampadumpara and Nedumkandam was similar to Banana bract mosaic virus (BBrMV) CdM isolate of Karnataka (91.01%), Coimbatore (90.29%) and Thrissur (95.76%) respectively. Phylogeny tree constructed in MEGA 6.0 software differentiated CdMV and BBrMV into four clades, in which CdMV Kattappana and Nedumkandam isolates were clustered together whereas CdMV Pampadumpara isolate was in separate clade. Similarly, BBrMV isolates of Pampadumpara and Nedumkandam clustered together while BBrMV Kattappana was in separate clade. Meristem of 2 mm size separated from infected plants were grown in Murashige and Skoog medium supplemented with 3 mg benzyl amino purine (BAP), 1.5 mg indole acetic acid (IAA) and 0.8 mg kinetin expressed direct organogenesis but multiple shoots were not produced. The TC plants were subjected to DAC-ELISA with the specific polyclonal antibodies and PCR with specific primers of the viruses and confirmed that the plants produced from meristems were free of both the viruses. Thus, the present study revealed that two viral diseases viz., katte and chlorotic streak affecting small cardamom in Idukki. Serologically and molecularly it was detected that katte disease was caused by Cardamom mosaic virus (CdMV) and chlorotic streak disease was caused by Banana bract mosaic virus (BBrMV), and the viruses could be eliminated from the infected plants through meristem tip culture to produce the diseases free plants.ThesisItem Open Access Process optimisation and quality evaluation of fruit pulp based yoghurts(Department of Community Science, College of Horticulture,Vellanikkara, 2019) Lovely Mariya, Johny; KAU; Seeja Thomachan, PanjikkaranYoghurt is a coagulated milk product that results from the fermentation of lactose in milk by Lactobacillus bulgaricus and Streptococcus thermophilus. Yoghurt is known for its nutritional, therapeutic and functional properties. The present study was undertaken to standardise fruit pulp based yoghurts and to enrich the standardised products with functional ingredients like garden cress seed and flax seed. The study also envisaged to evaluate the organoleptic, physico chemical, nutritional and keeping qualities of the standardised products. Fruit pulp based yoghurts (FPBY) were prepared using locally available fruits such as sapota, guava, jackfruit (Koozha type), banana (Palayamkodan) and papaya. Twenty six treatments were evaluated for various organoleptic qualities and was compared with 100 per cent milk yoghurt (control). In different treatments tried for the preparation of FPBY, the mean scores for different quality attributes showed a decreasing trend with increase in the quantity of fruit pulps. Yoghurts prepared by incorporating 10 per cent fruit pulp was selected as the most acceptable treatment from all FPBY. Physicochemical properties like moisture and pH decreased during storage and a gradual increase in acidity, syneresis, curd tension and viscosity was observed in FPBY. In the selected products TSS, reducing sugar, total sugar, energy, carbohydrate, lactose, fat, vitamin C and iron content decreased during storage. Protein content of freshly prepared control yoghurt was 4.59 per cent and on 15th day of storage it decreased to 3.35 per cent. The protein content varied from 3.76 to 2.71 per cent in SPBY (Sapota pulp based yoghurt), 3.23 to 2.61 per cent in GPBY (Guava pulp based yoghurt), 3.91 to 3.08 per cent in BPBY (Banana pulp based yoghurt), 4.42 to 3.22 per cent in JPBY (Jackfruit pulp based yoghurt) and 3.23 to 3.24 per cent in PPBY (Papaya pulp based yoghurt) in initial and at the end of storage respectively. Vitamin A and vitamin C content was found to be increased with incorporation of fruit pulp. The highest calcium content was found to be in control (78mg/100g) and a slight decrease in calcium content was observed among FPBY. A slight increase in iron and potassium content was observed in FPBY compared to control. The highest potassium content was observed in BPBY with an initial content of 83.25 to 83.05 mg/100g at the end of storage. The selected FPBY was incorporated with garden cress seeds (GCS) and flax seeds (FS) at 0.5 per cent and 2 per cent level, respectively. The highest total organoleptic scores were attained for control (51.67) which was incorporated with garden cress seeds. This was followed by GPBY (48.45), SPBY (48.33), JFPBY (47.81), BPBY (46.73) and PPBY (43.72). In flax seeds incorporated yoghurts the highest organoleptic score was attained for control (40.78) which was followed by GPBY (40.15), SPBY (40.14), BPBY (39.94), PPBY (39.82) and JFPBY (39.78). Incorporation of 0.5 per cent garden cress seeds and 2 per cent flax seeds increased the protein content from 4.59 per cent to 4.78 and 4.93 per cent respectively. Incorporation of functional ingredients increased the fat, iron, calcium and potassium content in all yoghurts. During storage, E coli and coliform bacteria were not detected in all yoghurt samples. Yeast and fungi was not detected up to 10th day of storage and at 15th day of storage the presence of yeast and fungi were observed among all the selected yoghurt samples, indicating a shelf life of ten days. The cost of production of plain yoghurt was Rs. 13.50/100 ml and for FPBY it varied from Rs. 17.00 to 20.00/100 ml. Incorporation of functional ingredients increased the cost of production from Rs. 17.16 to 21.00/100 ml. The standardised products are suitable for commercialisation and diversification. Milk based functional food products will be a boon to the dairy food industry.ThesisItem Open Access Process optimisation and quality evaluation of cocoa based chocolates(Department of Community Science, College of Horticulture,Vellanikkara, 2019) Shahanas, E; KAU; Seeja, Thomachan PanjikkaranCocoa (Theobroma cacao) is a supporting crop to farmers due to its remunerative income, which forms the basis for one of the world’s most popular food products chocolate. The quality of final product depends upon the fermented dried beans. High free fatty acid content is a serious quality defect and reduces the economic value of the cocoa beans. Recently, the cocoa trade has assumed a more scientific position and a lot of emphasis are placed on the content of free fatty acid. A reduction in the free fatty acids level will definitely have a positive impact. As such there is a need for the farmers to carry more intensive research and to develop and technology for the standardisation of free fatty acid content in the processing stages of cocoa in both primary and secondary processing. The present study was undertaken to develop protocol for primary processing of cocoa beans based on free fatty acid content (<1.75 %), to standardise the time and temperature of chocolate making using machine and to evaluate the quality attributes and shelf life of the products. Cocoa fermentation was carried out with three different types of fermentation methods like basket, heap and sack method for the periods of 5, 6 and 7 days of fermentation with nine different treatments. The physico-chemical qualities of fermented cocoa beans were evaluated. The highest (84.99%) fermentation index or cut test score was recorded in heap method at seventh day of fermentation (T6). The highest fermentation bean recovery of 84.33% was noticed in treatment T4 in heap method at fifth day of fermentation. The moisture content vary from 50.40 %, 53.67% and 54.40% to 39.80%, 37.83% and 39.43% from initial to seventh day of fermentation in sack, heap and basket method respectively. The pH of fermented beans varied from 6.11 to 4.26 in all methods. Among the fermentation methods the maximum fat content was in heap method from initial 42.66% to 32.89%. The lipase activity, decreased with fermentation periods in all three fermentation methods. In heap method it varied from 0.0022 to 0.0005. The heap method with seven days of fermentation were selected to sun drying and oven drying based on the lowest free fatty acid content (0.80%). The bean recovery of fermented dried beans was high (41.00 %) in sun dried cocoa beans as compared to oven dried beans (40.12 %). The free fatty acid content (1.26 %) in sun dried cocoa beans was less compared to 1.47% in T2 (oven dried beans). The moisture content (3.76%) and lipase activity (0.0018μ eq) was less but, the fat content and pH was high in sun dried beans. The sun drying proved to be adaptive as free fatty acid content was lower than oven dried beans and were below the cut off value of 1.75%. The sun dried beans were stored in gunny bags (control), polythene covers and plastic containers under ambient condition for a period of six months and the physico-chemical qualities of stored cocoa beans were done initially and at an interval of one month for a duration of six months. The highest bean recovery (98%) and least moisture content (4.22 to 4.52) was in cocoa beans stored in plastic container at the end of sixth month of storage. The lowest free fatty acid content of 1.68% was in cocoa beans stored in plastic container at sixth month after storage. The stored cocoa beans in plastic container were divided into two lots, one lot was used as such and the second lot was subjected to alkalisation. The chocolates was prepared with alkalised and non alkalised cocoa beans in tempering and conching machine for different time periods of 7, 9 and 11 hours at a temperature of 600C and 700. After the preparation of different types of alkalised and non alkalised chocolates in tempering and conching machine, chocolates were subjected to analysis of physico-chemical qualities, organoleptic evaluation and for enumeration of microflora. The treatment T6 (Chocolate prepared at 700C for 11 hours) and T12 (700C for 11hours) had the lowest free fatty acid content of 1.67% and 1.68% and maximum score for overall acceptability (8.89 and 8,87). Chocolates prepared at 700C for 11 hours from alkalised and non alkalised beans were blended with dehydrated grapes, dehydrated dates, osmodehydrated jackfruit, osmodehydrated pineapple, badam, cashew nut, powdered dehydrated mint leaves and white pepper powder. From the, organoleptic evaluation of different treatments, the best rated treatments from alkalised chocolates were T3 (95 % alkalised chocolates +5% osmodehydrated jack fruit), T6 (95 % alkalised chocolates +5% badam) and T8 (95 % alkalised chocolates +5% white pepper) and from non alkalised chocolates the treatments T11 (95 % non alkalised chocolates +5% osmodehydrated jack fruit), T14 (chocolate blended with 95 % non alkalised chocolates and 5% badam) and T16 (95 % non alkalised chocolates and 5% white pepper powder) were selected, packed in aluminium foil and stored in refrigerator for six months. Initially the lowest hardness of 74.41N and 76.71N was recorded in controls (T0 - AC) and (T0 – NAC. The maximum energy content of 580.15Kcal was found in treatment T5 (95 % NAC+5% B). Initially the maximum protein content (7.74 g/100g) was noticed in treatment T2 (95 % AC+5% B) and T5 (95 % NAC+5% B). The highest polyphenol content ranging from 0.23g/100g to 0.19g/100g was seen in T0 (100% NAC) throughout the storage periods. A reduction in mineral contents like calcium, phosphorus and iron content was observed in all treatments and at the end of sixth month calcium content varied from 0.39g/100g to 0.63g/100g. An increase in lipase activity and free fatty acid content of blended chocolates was recorded during storage and FFA was within 1.75% up to fourth month of storage. Based on organoleptic score, the treatment T2 (95 % AC+5% B) attained the highest rank. The blended chocolates were evaluated for the presence of bacteria, yeast and mould initially and monthly intervals for a period of six months, At the end of sixth month of storage the highest bacterial count of 2.0 x 104 cfu/g was found and during sixth month of storage. The cost for blended chocolates was varied from Rs.46.54 to Rs.50.62 / 40 g of chocolate bar. The cost of the chocolates prepared in the present study was lesser as compared to the commercial chocolates. The present study found that good quality, nutritious and healthy blended chocolates using cocoa could be prepared without adding any preservatives. The blended chocolates contain treasure of nutrients and bioactive compounds polyphenol which is essential for the growth and development of children and also give several health benefits.ThesisItem Open Access Morpho-Molecular charaterisation of intergeneric hybrids of Ascocentrum(Department of Floriculture and Landscaping, College of Horticulture, Vellanikkara, 2019) Katare Renuka, Shamrao; KAU; Sobhana, A.Morpho-molecular characterisation intergeneric hybrids of Ascocentrum was conducted at the Department of Floriculture and landscaping, College of Horticulture, Vellanikkara, during 2016-19 with the objective of characterising based on morphological and molecular analysis for commercial exploitation and compatibility assessment. Thirty varieties selected for the morphological characterisation. In quantitative characters Mok. Omayaiy Yellow showed highest plant height, internodal length and leaf breadth throughout the study period. Plant spread, leaf length and leaf area were highest in Kag. Youthong Beauty. Mok. Sayan × Ascda. Doung Porn was observed with highest shoot girth, shoot diameter and number of leaves. Ascda. Yip Sum Wah × V. JVB showed maximum number of roots, Mok. Khaw Phiak Suan × Ascda. Jiraprapa was observed with the highest root length and Mok. Chao Praya Sunset Yellow Spot with the highest root girth. Vasco. Aroonsri Beauty had the least values in all vegetative characters throughout the study period including plant height, spread, internodal length, shoot, leaf and root characters except the number of leaves which was the least in Mok. Khaw Phiak Suan × Ascda. Jiraprapa. Variety Vasco. Aroonsri Beauty also produced the maximum number of florets/spike and had the shortest internodal length. Cluster analysis with 14 different floral characters revealed 12 clusters at 75 per cent similarity. The highest inter-cluster distance was observed in cluster 6 and cluster 10. Cluster 6, which included Ascda. Sirichi Fragrance and Vasco Blue Bay White was found to have the lowest internodal length with the highest value LVII for number of florets per spike, also observed to have lower flower length and flower width. Cluster 10 was found to have the high mean values for spike length, flower length and flower width. In qualitative characters, based on growth habit, two types were found among the varieties viz., hanging and prostrate nature of growth. Leaf texture was found smooth and rigid with entire leaf margin. Leaf apex was acute in Vasco. Aroonsri Beauty and Ascda. Yip Sum Wah × V. JVB and was emarginated in rest of the hybrids. Wide variation was found among the flower colour, colour pattern, nature of petals and lip. Regarding post harvest traits, variety Mok. Omayaiy Yellow recorded highest fresh weight of spike and physiogical loss in weight. Whereas, variety Kag. Youthong Beauty took maximum days to start wilting of a floret. Mok. Chark Kuan Pink was observed to have longest vase life, spike longevity and highest water uptake. In visual evaluation the highest total mean score for the spike to use as a cut flower was observed in Mok. Omayaiy Yellow (54.6 out of 60), while the lowest was observed in Vasco. Blue Bay White. The highest mean score was obtained in Vasco. Pine River Pink (53.83) for plants for the indoor display. In pollen studies Ascda. Sirichi Fragrance and Mok. Sayan × Ascda. Doung Porn showed the highest pollen fertility and germination and these were selected as two male parents for further cross compatibility check. Vasco. Aroonsri Beauty and Kag. Youthong Beauty were found self-incompatible as well as cross incompatible with both the male patents and Vasco. Pine River Blue was found cross incompatible with Mok. Sayan. × Ascda. Doung Porn whereas, the rest of the varieties were found cross-compatible with both the male parents. LVIII Among the 21 SSR primers, ten generated polymorphic patterns. The number of amplicons detected varied from two to seven. The highest number of alleles was found in FJ539054, FJ539061 and JN375718. Primers DQ494847 (3) observed to have less number of amplicons. The PIC value ranged from 0.095 to 0.800.. One unique band was produced by JN375713 and FJ539050 primers in Kag. Samrong and Vasco. Aroonsri Beauty, respectively. The least Jaccard’s similarity value (0.05) was observed between Kag. Samrong and Ascda. Suksamran Sunlight, Ascda. Yip Sum Wah × V. Josephine Van Brero, Vasco. Aroonsri Beauty. The UPGMA clustering algorithm grouped the varieties into two main clusters. The variety Kag. Samrong clustered separately from all other members, whereas, other members were grouped in one cluster. At 50 per cent level of similarity, the hybrids grouped into 13 clusters. Out of 29 ISSR primers used, 20 showed amplification in all hybrids with polymorphic bands. ISSR primer (GACAC) 4 generated 11 amplicons, whereas, ISSR 901 generated 31 amplicons. ISSR primer (GACAC) 4 had lowest PIC value, and UBC810, the highest PIC value (0.926). The least Jaccard’s similarity value (0.03) was observed between Vasco. Blue Bay White and Mok. Khaw Phiak Suan × Ascda. Bicentennial Yellow Spot, which indicates that these hybrids are dissimilar to each other. The highest Jaccard’s similarity value was observed between Mok. Khaw Phiak Suan × Ascda. Bicentennial Yellow Spot and Mok. Chao Praya Sunset Yellow Spot. A UPGMA-based dendrogram separated the 20 hybrids of Ascocentrum orchids into two main clusters, each with 10 members. At 30 per cent level of similarity, all the 20 hybrids grouped into 14 different clusters. Six clusters were observed with two members each.ThesisItem Open Access Investigations on yellowing of black pepper (piper nigrum L.)(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2019) Maddirala Surendra, Babu; KAU; Sujatha, V SBlack pepper, popularly known as King of Spices or Black gold is one of the important and earliest known spices produced and exported from India. After foot rot disease, the major constraint in black pepper cultivation is the incidence of yellowing, leading to decline in production year after year. Yellowing is reported to be caused by many abiotic and biotic factors. Symptomatological studies on yellowing showed that per cent severity of yellowing was significantly higher during October-November and was on par during summer. During July-August, yellowing was significantly low. There was no definite pattern in the development and spread of the disease. In case of mild yellowing, there was recovery during July-August. Yellowing gradually increased and severely affected plants failed to recover. There was no flushing or flowering in plants. Such plants died in 2-3 years. All varieties studied were found to be susceptible to yellowing. Yellowing was observed either at the top of pepper column or (and) the bottom of the column. Sometimes yellowing appeared in a group of leaves together. In some aged plants older leaves at terminal portion of plant were yellow. In some leaves the base of lamina near petiole was yellow. This yellowing faded to tip of lamina and margins. Yield and yield contributing characters in healthy, apparently healthy and yellowing affected black pepper plants showed that all the characters of healthy plants showed a higher positive value compared to yellowing affected plants. In the case of yield per plant, there was significant reduction in yield in yellowing affected plants compared to healthy in all varieties except Vijay. In the study on rhizosphere soil nutrient status there was no apparent role of soil pH, OC, N, P, K, Ca, Mg, S, Fe, Mn, Zn, Cu and B in causing yellowing in the experimental plots in the present study. All the nutrient elements were in the optimum or high range. There was no significant difference among yellowing affected, apparently healthy and healthy plants in case of N and S where DRIS norms were not available. Among the rhizosphere soil parameters, pH, EC and organic carbon were significantly and positively correlated with yellowing. There was no significant correlation of N, P, Mg, S, Fe, Mn, Zn, Cu and B content of rhizosphere soil with yellowing in black pepper. K, Ca and Al content of rhizosphere soils showed significant negative correlation with yellowing. Analysis of nutrient status in the plant tissues of yellowing affected, apparently healthy and healthy plants based on DRIS norms suggested by Hamza et al. (2007) indicated that there was deficiency of N in the yellowing affected plants, K content and S content was also low in yellowing affected plants, all other nutrients were not deficient in the yellowing affected plants. Study on soil micro flora (cfu g-1) showed that mean value of fungal population was significantly highest (18.78 ×10 3cfu g-1) in rhizosphere soils of healthy plants compared to apparently healthy and yellowing affected plants. Healthy plants showed significantly low (27.55 ×10 3cfu g-1) population count of actinomycetes in the rhizosphere soil, whereas apparently healthy and yellowing affected plants were statistically on par. Yellowing affected plants showed significantly highest population count of bacteria (25.53 ×10 8cfu g-1) followed by apparently healthy. Healthy plants showed significantly lowest bacterial population. No soil borne pathogens including Phytophthora, Rhizoctonia or Fusarium was found associated with yellowing Nematode population in the rhizosphere soils of healthy plants was significantly lowest compared to apparently healthy and yellowing affected plants. Yellowing affected plants showed significantly highest population of Meloidogyne incognita, Radopholus similis, Pratylenchus sp, Helicotylenchus sp, Dorylaimid sp, and Trophotylenchulus sp, in the rhizosphere soil when compared with apparently healthy and healthy plants. In case of root, number of galls on the roots was significantly highest in yellowing affected plants. Considering different categories of nematodes, Meloidogyne incognita followed by Radopholus similis were predominant in the roots of yellowing affected plants. In roots as well as rhizospere soil, significantly highest population of nematodes was observed during October – November followed by February – March. Examination of roots of experimental plants and rhizosphere soil did not show the presence of root mealy bugs or any other insect, which can cause damage to root or yellowing in the plant. There was significant positive correlation of yellowing with maximum temperature. However minimum temperature showed significant negative correlation with yellowing. Relative humidity and rainfall also were significantly and negatively correlated with yellowing. A definite association of plant parasitic nematodes was seen in the plants as well as rhizosphere soils of yellowing affected plants in the present study. Meloidogyne incognita followed by Radopholous similis population was significantly highest in yellowing affected plants indicating their role clearly in causing yellowing. In the present study on the analysis of different factors like soil nutrients, plant pathogenic micro-organisms, soil borne insects, nematodes etc, in causing yellowing gives a clear indication of role of nematodes in causing yellowing in black pepper in the experimental plot in Thrissur district. Even though the nutrient status in the rhizosphere soil of experimental plants was satisfactory, absorption of nutrients was low due to damage of roots caused by nematodes. There was deficiency of nitrogen and low potassium and sulphur levels in the leaves of yellowing affected plants. The yellowing expressed must be due to multiple factors and combined effect of multiple nutrient deficiencies in plant tissue and damage due to nematodesThesisItem Open Access Performance analysis of medicinal kaempferia species(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2019) Akoijam Ranjita, Devi; KAU; Mini Raj, NThe medicinal Kaempferia species are a good source of valuable bioactive compounds. Kaempferia rotunda is widely used in the ancient systems of medicine in India and Indonesia. Kaempferia parviflora, popularly known as black ginger or Thai ginseng, has potential for great exploitation on commercial basis. Rhizomes of K. parviflora are used as aphrodisiac in traditional medicine in Thailand. The present study was undertaken at Department of Plantation crops and Spices, Kerala Agricultural University, Thrissur to evaluate the medicinal Kaempferia species in terms of morphology, anatomy, floral biology, yield, medicinal as well as pharmacological properties. A total of 18 genotypes belonging to three different species of Kaempferia formed the material for the study. Kaempferia galanga was taken as a reference species. The morphological evaluation was done consecutively for two years (2017-18 and 2018-19) based on qualitative and quantitative parameters. In K. rotunda, there was not much variation in qualitative parameters while the quantitative parameters exhibited tremendous variation among the 13 genotypes evaluated. The fresh rhizome yield ranged from 15.21 to 52.44 g/plant and dry yield of rhizome ranged from 5.00 to 17.73 g/plant. The Manipur collection MCR-6 performed best with the highest fresh as well as dry rhizome yield of 52.44 g and 17.73 g respectively followed by KCR-5 from Kerala. In K. parviflora also, the qualitative parameters did not show much variation whereas the quantitative characters exhibited significant variation among the genotypes. Significantly highest (114.60 g) fresh weight of rhizome was recorded by the genotype KCP-1. The Thailand collection KCP-1 was the best performer followed by BSI-1 from Shillong. The two genotypes of K. galanga evaluated in the study exhibited morphological variation in certain characters, however no significant variation was noticed for rhizome yield. The flowering was observed from May to November in K. parviflora, March to April in K. rotunda and June to July in K. galanga. In the floral biology studies, variation was noticed in floral parts including the pollen grains of the three species with respect to size and shape. The flowers were bisexual, complete, trimerous and zygomorphic in all the species. There were on an average 14.2 number of flowers per inflorescence in K. parviflora, 8.9 flowers in K. rotunda and 5.6 flowers per inflorescence in K. galanga. The time of anthesis was 5.00 am to 7.15 am in K. parviflora, 4.00 to 5.00 am in K. rotunda and in case of K. galanga, peak anthesis time was 4.00 to 5.00 am. The stigma was receptive upto eight hours after the anthesis in K. parviflora whereas K. rotunda and K. galanga remained receptive for 24 hours and nine hours respectively after anthesis. In the in vivo pollen germination studies, pollen germination was noticed only in K. parviflora and K. galanga. The seed set was observed only in K. parviflora. The vivipary was observed in K. parviflora and recalcitrance of the seed was confirmed, thus proving its viviparous nature. The viviparous plants were compared with rhizome borne plants. The number of leaves and tillers were more in viviparous plants when compared with normal plants but they took two years for appreciable yield. However, rhizome yield of viviparous plant was only 1/3rd of that in normal plants. In the anatomical studies, all the three species had collateral and closed type of vascular bundles in transverse section of leaves. The oil globules were present in the leaf lamina of K. galanga. Calcium oxalate crystals were present in the leaf epidermis of K. rotunda and leaf lamina in K. parviflora. Oil globules were abundant in the rhizome of K. rotunda and K. parviflora. Starch granules in rhizome and root tuber were concentrated near endodermal layer in all the species. The flavonoid vacuoles were abundantly present in the rhizome section of K. parviflora. In the biochemical studies, the volatile oil content in K. rotunda rhizome ranged from 0.057 to 3.17 per cent and oleoresin content was to the tune of 0.60 to 3.17 per cent. In K. parviflora, volatile oil content was negligible; oleoresin content ranged 2.03 to 4.17 per cent. The content of starch, total sugars and flavonoids were high in K. parviflora whereas total free amino acid content was high in K. rotunda rhizome. The profiling of volatile oil of K. rotunda by GCMSMS detected 22 compounds, that of K. parviflora indicated 34 compounds and in K. galanga there were 27 compounds. The profiling of ethanolic extract of K. rotunda rhizome by GCMS showed the presence of 18 compounds and that of K. parviflora, eight compounds. The ethanolic extract of rhizome of K. rotunda and K. parviflora were subjected to detailed in vitro as well as in vivo pharmacological studies at the Dept. of Pharmacology and Toxicology, KVASU, Mannuthy. The in vivo acute toxicity and immunomodulatory study was carried out in Swiss albino mice using cyclophosphamide as immunosuppressive agent. No acute toxicity was noticed in the ethanolic extract of both K. rotunda and K. parviflora. Both the species significantly increased the body weight, total leukocyte count, serum protein and decreased neutrophil count in normal as well as immunosuppressed animals. A significant stimulation of humoral and cellular immune response was indicated by increase in antibody titre, bone marrow cellularity and DTH reaction. Histopathology of spleen confirmed the high immunomodulatory effect of K. parviflora and moderate immunomodulatory activity of K. rotunda. In the DPPH assay, K. rotunda exhibited high antioxidant activity (IC50 131.15 μg/ml) while K. parviflora showed lower activity (IC50 198.68±7.62 μg/ml). Ethanolic rhizome extract of both the species exhibited anticancer property in breast cancer cell lines. Both K. rotunda and K. parviflora showed cytotoxicity against MDA MB231 and MCF-7 cell lines. IC50 for K. rotunda was 167.1±5.60 and 194.8±8.97 respectively for MCF-7 and MDA MB231 cell lines while for K. parviflora it was 143.03±2.70 and 126.35±2.53 respectively. The ethanolic extract of K. rotunda and K. parviflora exhibited appreciable antimicrobial activity on E. coli (11.13±0.16 mm, 12.32±0.12 mm), S. enterica (11.47±0.29 mm, 13.8±0.16 mm) and P. aeruginosa (11.52±0.38 mm, 11.17±0.31 mm) and showed potent activity on S. aureus (14.18±0.32 mm, 15.48±0.23 mm).
