PG Thesis
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Item Diversity analysis of chittamruthu [Tinospora cordifolia (WILLD.)] accessions from the northern districts of Kerala(Department of Plantation Crops and Spices, College of Agriculture, Vellayani, 2023-02-10) Dhanya, T; Sonia, N SThe present investigation entitled “Diversity analysis of Chittamruthu [Tinospora cordifolia (Willd.)] accessions from the Northern districts of Kerala” was carried out in the Department of Plantation Crops and Spices, College of Agriculture, Vellayani during the period 2020-2022 with the objectives to estimate the genetic variation in Chittamruthu accessions using SSR markers and its phytochemical screening. Chittamruthu plant samples (25 nos.) were identified and collected from 25 different locations in the 13 Agro-Ecological Units (AEUs) of North Kerala viz., AEU 2 [Mundayad, Kannur and Elathur, Kozhikode], AEU 6 [Thavanoor, Malapuram and Iringalakkuda, Thrissur], AEU 7 [Cherukunnu, Kannur and Pilicode, Kasaragod], AEU 10 [Chelakkara, Thrissur and Madakkathara, Thrissur], AEU 11 [Kakkodi, Kozhikode and Elamkulam, Malappuram], AEU 13 [Nilamboor, Malapuram and Peravoor, Kannur], AEU 15 [Kelakam, Kannur and Kottiyoor, Kannur], AEU 18 [Attapadi, Palakkad and Sholayur, Palakkad], AEU 19 [Attapadi, Palakkad and Pudur, Palakkad], AEU 20 [Ambalavayal, Wayanad and Kalpatta, Wayanad], AEU 21 [Pulpalli, Wayanad and Thirunelli, Wayanad], AEU 22 [Puthuppariyaram, Palakkad and Vadakkancherry, Palakkad] and AEU 23 [Pattancherry, Palakkad]. The indigenous traditional uses of Chittamruthu plants documented through a survey done in these locations are as follows. The plant could cure cough and cold, acts as an anti-arthritis agent, a remedy for headache, treatment for indigestion and constipation, cure skin issues, cure fever, relieve pain and inflammation in joints, cure for urinary infection, treating diabetes, remedy for piles, control complications of dengue fever, control the risk of cardio- vascular disorders and used as a liver tonic. Morphological diversity analysis of Chittamruthu accessions had identified a superior accession from North Kerala viz., accession 1 (T1) from Mundayad- Kannur (AEU 2) possessing significantly higher stem diameter (1.93 cm), density of lenticels (141.33 no.s), leaf petiole length (11.13 cm), laminar length (11.07 cm), laminar breadth (12.03 cm) and leaf area (117.67 cm2 ). Molecular diversity analysis of Chittamruthu accessions were carried out using 10 SSR primers. Among them, four primers viz., TCTSSR 104, TCTSSR 92, TCTSSR126 and TCTSSR-59 found to be polymorphic in nature. The primer TCTSSR 104 was found to be more informative for diversity analysis since, it has got higher polymorphism information content (PIC) value, 0.46. The dendrogram obtained from polymorphic SSR markers using NTSYS-pc software had grouped the 25 Chittamruthu accessions into four major clusters. Cluster I and II included two accessions, cluster Ⅲ contained three accessions and cluster Ⅳ was the largest cluster containing total of 18 accessions. There exists phytochemical diversity among the 25 Chittamruthu accessions. Qualitative analysis of Chittamruthu accessions exhibited the presence of alkaloids, flavonoids, phenols, saponins and cardiac glycosides. Quantitative phytochemical analysis revealed that T3, the accession obtained from Thavanoor, Malappuram (AEU 6) recorded the highest total alkaloid (102.89 µg AE mg-1 ), total flavonoid (54.29 µg QE mg-1 ), total phenol content (116.54 µg GAE mg-1 ) and cardiac glycosides (25.87 µg DE mg-1 ). Total saponins were found to be high (72.84 µg DE mg -1 ) in the accession, T13 obtained from Kelakam, Kannur (AEU 15). The study revealed that Chittamruthu accessions collected from 13 different AEUs of Northern districts of Kerala were found to be diverse in morphological, molecular and phytochemical characteristics. The accession collected from Mundayad, Kannur (AEU 2) found to possess superior morphological parameters and accession collected from Thavanoor, Malappuram (AEU 6) found to be superior in phytochemical quality. In addition, the Chittamruthu accessions collected from different AEU’s of Northern Kerala could be clustered into four main clusters based on molecular diversity.Item Diversity analysis of Chittamruthu [Tinospora cordifolia (wild)] ecotypes of southern districts of Kerala(Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture , Vellayani, 2024) Revathy, U.S.; Sonia, N.S.The study titled "Diversity analysis of Chittamruthu [Tinospora cordifolia (Willd.)] ecotypes of Southern districts of Kerala" was conducted at the Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture, Vellayani during 2021-2023. The study aimed to assess morphological, phytochemical and genetic variation in Chittamruthu using SSR markers. Chittamruthu ecotypes, 27 nos. were identified from different locations in the ten distinct Agro-Ecological Units (AEUs) of Southern Kerala viz., AEU 1 (Chirayinkeezhu, Kottiyam, Kundara), AEU 3 (Suranad), AEU 4 (Thiruvalla, Ambalappuzha), AEU 5 (Mala), AEU 8 (Neyyattinkara, Kalliyoor, Kattakkada), AEU 9 (Ponkunnam, Mylom, Vembayam, Pala), AEU 12 (Vannapuram, Nanniyod, Vellanad), AEU 14 (Kulathuppuzha, Nemmara, Vellathooval, Nelliyambathi, Adimali, Vithura), AEU 16 (Kattappana, Pambadumpara, Shanthanpara) and AEU 17 (Kanthalloor). Chittamruthu stem is being used by the localites for curing kidney-related ailments in Chirayinkeezh, Thiruvananthapuram. Chittamruthu stem powder could improve digestion as opined by the localites in Kottiyam, Kollam. Amrithasatwa is a unique Chittamruthu preparation revealed by a traditional medicinal practitioner in Suranad, Kollam for treating acidity. In addition, local use of Chittamruthu for addressing health issues viz., menstrual irregularities, post-partum care, managing rheumatic fever, diabetes, joint pain, arthritis and dengue fever were documented. Chittamruthu ecotypes showed significant morphological variations in stem diameter, density of lenticels, petiole length, leaf length, leaf breadth and leaf area. Ecotype (T6) collected from Ambalappuzha (AEU 4) recorded the highest stem diameter (2.00 cm), petiole length (12.60 cm), leaf area (153.78 cm2), laminar length (12.37 cm) and laminar breadth (12.67 cm). The highest density of lenticels was observed in the ecotype (T14) obtained from Pala (AEU 9). Genetic variation among the 27 Chittamruthu ecotypes were assessed using ten SSR markers viz., TCTSSR 59, TCTSSR 104, TCTSSR 126, TCTSSR 92, TCTSSR 37, TCTSSR 18, TC 23, TC 8, TC 9, and TC 1. Among these primers, TCTSSR 104, TCTSSR 92, TC 23, TCTSSR 59, TCTSSR 126 and TC 1 exhibited polymorphism, while the remaining four were monomorphic. TCTSSR 104 recorded the highest Polymorphism Information Content (PIC) value (0.49). A dendrogram developed using NTSYS-pc software demarcated the 27 Chittamruthu ecotypes into 96 seven clusters at a similarity coefficient, 0.86. Cluster I comprised of 20 ecotypes, Cluster II to VI were comprised of one ecotype each and Cluster VII comprised of two ecotypes. A single representative ecotype from each cluster was used for phytochemical diversity analysis. Vellanad ecotype (AEU 12) recorded the highest total alkaloid (183.96 μg AE mg -1) and total flavonoid (162.62 μg QE mg-1) content. The highest total phenol content (55.21 μg GAE mg -1), total saponins (99.29 μg DE mg-1) and cardiac glycosides (42.09 μg DE mg -1) were recorded by Chirayinkeezhu (AEU 1), Kulathuppuzha (AEU 14) and Nemmara (AEU 14) ecotypes, respectively. The study revealed that 27 Chittamruthu ecotypes collected from ten distinct AEUs from the Southern districts of Kerala showed significant diversity in their morphological, molecular and phytochemical characteristics. Ambalappuzha ecotype (AEU 4) recorded superior morphological characteristics. Molecular diversity analysis using SSR markers had demarcated the 27 Chittamruthu ecotypes into seven genetically diverse clusters. Pharmacological properties of Chittamruthu are mainly attributed to alkaloids. In this study, Vellanad ecotype (AEU 12) is identified as potential alkaloid yielding Chittamruthu (183.96 μg AE mg -1).