<Emphasis Type="Italic">Agrobacterium-</Emphasis>mediated genetic transformation of tea leaf explants: effects of counteracting bactericidity of leaf polyphenols without loss of bacterial virulence

Tea is one of the major crops in Asia and Africa, and its improvement by genetic modification is important for economy of many tea-producing regions. Although somatic embryos derived from cotyledon explants have been transformed with Agrobacterium, the leaves of several commercially important tea cultivars have remained recalcitrant to transformation, largely due to bactericidal effect of polyphenols that are exuded by tea leaves in vitro. Moreover, the commonly used polyphenol adsorbents and antioxidants cannot overcome this problem. Leaf explants, however, are more desirable than cotyledon-derived somatic embryos, especially when it is necessary to further improve a selected elite and also retain its superior traits. Thus, we developed a procedure for Agrobacterium-mediated genetic transformation of tea leaf explants which is based on the presence of l-glutamine in the co-cultivation medium. We then showed that the transformation process is facilitated via a protective action of l-glutamine against bactericidal effects of leaf polyphenols without affecting the bacterial virulence (vir) gene expression.     

Ectopic expression of SOD and APX genes in Arabidopsis alters metabolic pools and genes related to secondary cell wall cellulose biosynthesis and improve salt tolerance

Molecular Biology Reports - Hydrogen peroxide (H2O2) is known to accumulate in plants during abiotic stress conditions and also acts as a signalling molecule. In this study, Arabidopsis thaliana...     

Identification and cross-species amplification of EST derived SSR markers in different bamboo species

The availability of expressed sequence data derived from gene discovery programs became an alternative source of mining simple sequence repeat (SSR) and developing inexpensive genetic markers for the crop improvements. In present study, 10 express sequence tags (EST)-SSR markers were identified from Bambusa oldhamii public sequence data base. Transferability to 25 species of Bambusoideae ranged from 30% to 100%. The number of alleles detected per locus ranged from 2 to 10. All the newly identified SSR markers were found to be moderately to highly polymorphic with an average Polymorphic Information Content (PIC) value of 0.54. As these loci represents transcribed region and recorded high level of cross transferability and reliable amplification across the species, demonstrating the utility of these markers for functional and genetic analyses of bamboo species.     

Establishment and multiplication of colchi-autotetraploids of Rauvolfia serpentina L. Benth. ex Kurz. through tissue culture

A tissue culture procedure was developed for the establishment and propagation of a colchi-autotetraploid of Rauvolfia serpentina for possible commercial exploitation. Multiplication of autotetraploid shoots was obtained either through axillary bud elongation on Murashige and Skoog [1] medium (MS) containing 2.65 M (0.5 mgl^–1) -naphthaleneacetic acid and 0.33 M (0.05 mgl^–1) kinetin, or via multiple shoot formation on MS medium supplemented with 4.44 M (1.0 mgl^–1) 6-benzylaminopurine and 0.53 M (0.1 mgl^–1) -naphthaleneacetic acid. Rooting could be induced by transferring the shoots to MS medium containing 7.95 M (1.5 mgl^–1) -naphthaleneacetic acid alone. The plantlets, thus formed, were tetraploid in nature by cytological observations of the root tips. They exhibited 80–90% success in establishment under glass house and field conditions.     

An efficient liquid culture system for tea shoot proliferation

The efficiency of thidiazuron in promoting tea shoot proliferation in liquid medium was evaluated. As compared to 6-benzyl adenine which induced hyperhydricity in the proliferated shoots in liquid medium, a progressive increase in the multiplication rate together with healthy shoot growth was achieved when thidiazuron (2.5 to 5.0 μM) was used instead of 6-benzyl adenine. Of the different liquid volumes compared in 250 ml Erlenmyer flasks, 20 ml was the most effective. While an increase in the multiplication rate coupled with normal but healthy shoots was observed under static and agitated conditions at this volume of liquid medium, hyperhydricity was induced in 50 ml liquid medium. Therefore, 20 ml static liquid medium with subculture periods at an interval six to eight weeks seems to be a cost and labour effective process as compared to the existing protocols involving solid media with subculture periods at 4 weeks interval. This revised version was published online in June 2006 with corrections to the Cover Date.     

Two rounds of whole genome duplication in the ancestral vertebrate

The hypothesis that the relatively large and complex vertebrate genome was created by two ancient, whole genome duplications has been hotly debated, but remains unresolved. We reconstructed the evolutionary relationships of all gene families from the complete gene sets of a tunicate, fish, mouse, and human, and then determined when each gene duplicated relative to the evolutionary tree of the organisms. We confirmed the results of earlier studies that there remains little signal of these events in numbers of duplicated genes, gene tree topology, or the number of genes per multigene family. However, when we plotted the genomic map positions of only the subset of paralogous genes that were duplicated prior to the fish–tetrapod split, their global physical organization provides unmistakable evidence of two distinct genome duplication events early in vertebrate evolution indicated by clear patterns of four-way paralogous regions covering a large part of the human genome. Our results highlight the potential for these large-scale genomic events to have driven the evolutionary success of the vertebrate lineage.     

Do leaf surface characteristics affect<Emphasis Type="Italic">Agrobacterium</Emphasis> infection in tea [<Emphasis Type="Italic">Camellia sinensis</Emphasis> (L.) O Kuntze]?

The host range specificity ofAgrobacterium with five tea cultivars and an unrelated species (Artemisia parviflora) having extreme surface characteristics was evaluated in the present study. The degree ofAgrobacterium infection in the five cultivars of tea was affected by leaf wetness, micro-morphology and surface chemistry. Wettable leaf surfaces of TV1, Upasi-9 andKangra jat showed higher rate (75%) ofAgrobacterium infection compared to Upasi-10 and ST-449, whereas non-wettable leaves ofA. parviflora showed minimum (25%) infection. This indicated that the leaves with glabrous surface having lower 8 (larger surface area covered by water droplet), higher phenol and wax content were more suitable forAgrobacterium infection. Caffeine fraction of tea promotedAgrobacterium infection even in leaves poor in wax (Upasi-10), whereas caffeine-free wax inhibited bothAgrobacterium growth and infection. Thus, study suggests the importance of leaf surface features in influencing theAgrobacterium infection in tea leaf explants. Our study also provides a basis for the screening of a clone/cultivar of a particular species most suitable forAgrobacterium infection the first step inAgrobacterium-mediated genetic transformation.