Distant hybridization and compatibility studies in wild orchids

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Date
2018
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Department of Plant Breeding and Genetics, College of Agriculture, Vellayani
Abstract
The present investigation entitled “Distant hybridization and compatibility studies in wild orchids” was carried out with the objective of studying the cross compatibility through distant hybridization in wild orchids and molecular characterization of hybrid seedlings using RAPD/ISSR- markers. The study was carried out at Saraswathy Thangavelu Extension Centre of KSCSTE- JNTBGRI, Puthenthope, Thiruvananthapuram and at College of Agriculture, Vellayan, Thiruvananthapuram. The research work included experiment I, II, III and IV. Experiment I was carried out in two parts as evaluation of parents and hybridization, experiment II was cross compatibility studies of selected species, experiment III was in vitro culture of resultant hybrids and evaluation of seedlings and experiment IV was molecular characterization of hybrids. In experiment I, wild types belonging to different genera/species namely Vanda (Vanda tessellata, Vanda spathulata, Vanda sp.), Rhynchostylis retusa, Spathoglottis (Spathiglottis albida, Spathoglottis unguiculata, Spathoglottis gracilis, Spathoglottis plicata), Aerides ringens and Arundina graminifolia available at the orchid conservatory of Saraswathy Thangavelu Extension Centre of JNTBGRI, Puthenthope, Thiruvananthapuram were selected as parents based on variability in floral characteristics and these were evaluated for various quantitative and qualitative characters. Statistical analysis was carried out in Completely Randomized Design and analysis of variance revealed highly significant differences for all the biometrical characters included in the evaluation. Estimation of components of variance revealed that Phenotypic Coefficient of Variation (PCV) was higher than both Genotypic Coefficient of Variation (GCV) and Environmental Coefficient of Variation (ECV) for all the traits studied. PCV (143.93) and GCV (128.52) were the highest for number of spikes plant-1. Heritability was the highest for flower width (99.06 %) and it was high for all thirteen traits. Genetic advance as percent of mean was high for all the traits studied. It was the highest for number of spikes plant-1 (239.80). Estimated phenotypic correlation coefficient revealed highly significant and positive association between flower length and flower width (0.9519). The highly significant and negative association was obtained between internodal length and days for inflorescence emergence to first flower opening (-0.6896). Similarly genotypic correlation coefficient exhibited highly significant and positive association between flower length and flower width (0.9545) and highly significant and negative association between plant spread and number of leaves plant-1 (-0.9903). Based on this, parents were selected and were crossed in all possible combinations. In Vanda a hybrid evolved from the wild species was available at the centre and it was also crossed with the wild parents to study the cross compatibility of wild species with hybrids. In experiment II, cross compatibility of 152 cross combinations made was studied based on successful capsule setting. Capsule characteristics were recorded. The genus Spathoglottis took only four days for attaining pod setting whereas for the genus Rhynchostylis it was ten days for pod setting. Capsule attained maturity much earlier (30 days) in the genus Spathoglottis but the species Vanda spathulata took more days for attaining capsule maturity i.e., harvesting stage (150-154 days). In experiment III, capsules developed among thirty one cross combinations were subjected to seed culture for rapid germination under in vitro condition and seedling development. Observations regarding seed germination were recorded and seeds of ten crosses exhibited seed greening and initiation of protocorm development. Seeds of seven cross namely one intergeneric hybrid and six interspecific hybrids were developed into seedlings. Evaluation of hybrid seedlings was made based on variability in morphological characters at monthly intervals. The six interspecific hybrids were Spathoglottis unguiculata x Spathoglottis albida (Culture 622), Spathoglottis gracilis Spathoglottis unguiculata (Culture 652), Spathoglottis albida x Spathoglottis unguiculata (Culture 778), Spathoglottis albida x Spathoglottis unguiculata (Culture 1368), Spathoglottis plicata x Spathoglottis unguiculata (Culture 1386), Spathoglottis unguiculata x Spathoglottis plicata (Culture 1388) and intergeneric hybrid was Spathoglottis unguiculata x Vanda sp. (Culture 679). In experiment IV, molecular characterisation of seedlings evolved through the crossing was done based on ISSR analysis to distinguish F1 hybrid seedlings from selfed seedlings and their parents. Percentage of polymorphism between hybrids and parents ranged from 11.99 to 32.97 and these results confirmed that all the seven crossed seedlings tested were F1 hybrids and these were distinct from their relevant parents. The above study revealed that through distant hybridization in wild orchids, one intergeneric hybrid Spathoglottis unguiculata x Vanda sp. and six interspecific hybrids viz., Spathoglottis unguiculata x Spathoglottis albida, Spathoglottis gracilis x Spathoglottis unguiculata, Spathoglottis albida x Spathoglottis unguiculata, Spathoglottis albida x Spathoglottis unguiculata, Spathoglottis plicata x Spathoglottis unguiculata, Spathoglottis unguiculata x Spathoglottis plicata could be obtained and molecular characterization confirmed the hybridity of the intergeneric and interspecific hybrids. Therefore, the intergeneric and six interspecific F1 hybrids evolved through distant hybridization can be carried forward for the development of varieties or can be utilised for genetic improvement programme in future.
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