Kadanthottu Sebastian Joseph

Development of SCAR marker for sex identification in dioecious Garcinia gummi-gutta

Key messageWe have developed sex-specific SCAR marker for the identification of dioeciousGarcinia gummi-gutta(L.), which is useful for the selection ofG. gummi-guttaat seedling stage and for plantation programmes.AbstractGarcinia gummi-gutta (L.) Robs. is a dioecious fruit yielding tree, which is naturally distributed as well as cultivated in the orchards in Western Ghat regions of India. A sex-linked DNA fragment was identified in Garcinia gummi-gutta (L.) Robs. by screening 150 randomly amplified polymorphic DNA primers and only one of them (OPBD20) showed different amplification band pattern associated with sex type. This sex-linked fragment was converted into male-specific sequence-characterized amplified region (SCAR) marker, CAM-566. The primers deigned in this study (OPBD20F and OPBD20R) correctly differentiated 12 male and 12 female plants at high annealing temperatures. Thus, a 556-bp band was amplified in male samples but not in female ones. Nevertheless, it should be noted that the fragments from both sexes were amplified at relatively low annealing temperatures. Additionally, the developed SCAR marker successfully identified the sexes of ten sex-unknown samples. Therefore, it can be used as an effective, convenient and reliable tool for sex determination in such dioecious species.

Development of male-specific SCAR marker in Garcinia morella (Gaertn.) Desr.

Garcinia morella (Gaertn.) Desr. (family Clusiaceae) is popularly known as ‘Indian gamboge’ is a fruit yielding tree of tropical rain forests of Western Ghats of India (Anonymous 1956). It also occurs in Sri Lanka and Indo-China Himalayan regions. It is a multipurpose tree grown as a plantation crop along with Garcinia indica (Kokum) and Garcinia cambogia (Malabar tamarind). The fruit rinds are used as a condiment and for garnish. Bioactive compounds such as moreollin (Subba Rao et al. 1978), gambogic acid (Tang et al. 2011) have been isolated from the fruits and the bark, respectively, and evaluated for their antibiotic and anticancer properties. The trees are dioecious and the distinction between male and female trees can be made only at the flowering stage, after 10–12 years. It is cultivated as a plantation crop and the sex determination in this plant at an early juvenile stage will be useful for planning the male and female tree ratio in the orchards and it also enables the tree improvement programme. Recently, molecular tools were employed in dioecious taxa for early identification of sex and understanding the developmental and the evolutionary pathways of sexual dimorphism. Specific molecular markers can be deduced from unique, single-copy segments of the genome and can be considered codominant and can be used in sex determination. Sequence characterized markers (SCAR) which are based on randomly amplified polymorphic DNA (RAPD) analysis are locus-specific, more reliable and more reproducible for molecular identification (Paran and Michelmore 1993). SCAR marker linked to sex-specific genes have beensuccessfully used in sex identification of many dioecious plants including Carica papaya (Bedoya and Nunez 2007), Phoenix dactylifera (Dhawan et al. 2013). In this

In Vitro Propagation of Artemisia japonica

ABSTRACT An effective protocol for in vitro propagation was developed for Artemisia japonica using nodal explants. The efficiency of the explants to induce the multiple shoots on Murashige and Skoog (MS) medium supplemented with 2.5, 5.0, 7.5, and 10 µM of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1-napthalene acetic acid (NAA), 2,4-dichlorophenoxy acetic acid (2,4-D), 6-benzylaminopurine (BAP), thidiazuron (TDZ), kinetin (Kn), and 2-isopentenyl adenine (2iP) was tested. A maximum of 20 axillary shoots per explant were induced on MS medium supplemented with 10 µM BAP. Shoots were rooted on MS medium supplemented with IBA (10 µM). The resulting plantlets were hardened in a mixture of cocopeat and perlite (1:1). Cytological analysis revealed that regenerated plants had 2n = 18 chromosomes, which was same as that of the mother plant.

Development of a SCAR marker associated with male sex determination in Garcinia indica Choisy

Summary Garcinia indica is an endemic fruit-yielding tree which is naturally distributed in the Western Ghats regions of India. It is also cultivated as a plantation crop. It is dioecious tree with a long vegetative phase (10 - 12 years). There are no morphological or biochemical markers to differentiate the sex of individual trees before they reach maturity. In the present study, a randomly amplified polymorphic DNA (RAPD) marker was identified, using a pooled DNA approach, which was able to identify male trees. PCR screening of bulked DNA samples from male and female trees, using 150 decamer primers, revealed one primer (OPL-05) that produced a unique band of 1,501 bp in male tree DNA samples only. OPL-05 was then used to screen individual male and female trees in different collections. The 1.5 kbp band was produced in all male samples, but was absent in all female samples. This male-specific PCR fragment was cloned and sequenced and sequence-characterised amplified region (SCAR) primers were designed from the sequence of this unique fragment.The SCAR primers produced a 1,501 bp fragment in all male tree samples, but not in any female tree DNA sample.The SCAR primers were also used to investigate the sex of unknown trees before maturity.

Plant Cell and Organ Culture as an Alternative for the Production of Anticancer Compounds

Plants are the reservoirs of various valuable secondary metabolites like camptothecin, podophyllotoxin, and ginseng saponins which are used as anticancer agents. Plant cell and organ cultures are competent to synthesize and accumulate many of these compounds and can be used as a source for extraction of such compounds. Different strategies have been applied for overaccumulation of secondary metabolites in plant cell cultures which include screening and selection of high yielding cell lines, optimization of nutrient media, and elicitation. In the present chapter, some of the anticancer compounds derived from plant cell and organ cultures are described, and various strategies for improving the accumulation of useful secondary metabolites in cell and organ cultures and methods for the large-scale production using bioreactors are highlighted.

Phenolic content, antioxidant and antibacterial activities of Plectranthus rotundifolius tuber extracts

ABSTRACT The phenolic and flavonoid concentration and antioxidant and antibacterial activities of Plectranthus rotundifolius tuber extracts in petroleum ether, chloroform, and methanol were determined. The methanol extracts had the highest recovery and demonstrated the highest antioxidant potential and efficacy against the clinical bacterial strains tested as evident from the minimum inhibitory concentrations.

Physicochemical Characteristics of the Seed and Seed Oil of the Potentially Medicinal Plant Ziziphus oenoplia

ABSTRACT The proximate composition of seeds, physicochemical characteristics, and fatty acid profiles of Ziziphus oenoplia seed oil were determined in this study. The seeds possessed low moisture (4.54%) and high carbohydrate (42.96%) and protein content (40%), making the seed oil suitable for storage and consumption. The saponification value (197.80) of the seed oil makes it a promising source for the soap and shampoo industry. The iodine and saponification values are comparable to those of major edible seed oils such as groundnut and soybean. The high amount of monounsaturated fatty acids (53.41%), especially oleic acid (53.38%), present in the oil makes it a better source for a low-fat diet and may reduce the risk of various heart-related diseases.