The present research work reports the cytomorphological evaluation and an efficient protocol for in vitro micropropagation of an endemic, endangered and threatened herb Centella asiatica (L.) Urban (Mandukparni) from Himachal Pradesh, India. Plant species is a potent memory enhancer, antidiabetic, antioxidant, antimutagenic, anticancerous and also reported to have cardiovascular properties and used to cure chronic hepatitis disorders. Eight morphometric characters for each 10 different accessions were extensively studied, but no new morphotype was reported. As per the cytology, the present report (2n = 18) confirmed the earlier chromosome counts from India and abroad. As there is an important need to preserve this plant species and to make it available all over the year to pharmaceutical industries without causing loss of germplasm from the wild region, efficient in vitro micropropagation protocol through nodal explant has been developed. As per the results, the highest percentage of multiple shoot induction was 90.20 showing average 16.3 number of shoots on the medium augmented with 2.0 mg/l BAP + 0.5 mg/l Kn. Whereas, the combined concentration of 1.0 mg/l NAA + 1.0 mg/l IBA showed highest 92.2% root induction with average 16.5 number of roots per shoot. The survival rate of these plantlets under green house condition was 80%. This protocol can be used for further regeneration and genetic transformation studies in Centella
SummaryWe present a cytological study of species of the genus Chaerophyllum L. (Apiaceae) by analysing five accessions covering three species from the Western Himalayas, India. The species are collected and studied for detailed male meiosis, multiple associations and pollen fertility. All the studied species exist at diploid level (based on x=11), and show a meiotic chromosome count of 2n=22 at diakinesis and metaphase-I, and regular chromosome distributions at anaphases-I/II. C. aromaticum is reported for the first time from India and found to be in conformity with the earlier reports of 2n=22 from outside India. Out of three studied species, two (C. acuminatum and C. aromaticum) showed the presence of multiple chromosomal associations at metaphase-I of meiosis-I. The paper herein discusses for the first time the occurrence of multiple associations in these species. The course of meiosis in all studied species shows impaired meiotic course in the form of cytomixis, chromatin stickiness, laggards and chromatin bridges. Abnormal microsporogenesis results in heterogeneous-sized pollen grains and reduced pollen fertility.
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