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Kiwifruit) Plant Indian Journal of Biotechnology Vol 3, April 2004, pp 249-257 Biotechnological interventions for genetic amelioration of Actinidia deliciosa var. deliciosa (kiwifruit) plant DR Sharma* and Poonam Shirkot Department of Biotechnology, Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan 173230, India Kiwifruit (Actinidia deliciosa var deliciosa) is a recent introduction among the promising fruit crops. Popularity of its nutritious fruit berries has created a substantial demand for quality planting material, which needs to be planted in certain proportions of female and male plants for Actinidia is dioecious. The large-scale propagation of planting material can be achieved through micropropagation; whereas, it's quality and clonal nature need to be ensured, preferably, through molecular markers. With further popularity of this fruit crop, tailor-made genotypes will be required, which can be made possible through biotechnological interventions. As A. deliciosa is limited in its adaptation to a very narrow range of agro- climatic conditions, there is also a need to develop cultivars endowed with traits of wider adaptation. So that this fruit crop can be cultivated in larger range of geographical areas. Development of micropropagation protocols, identification of gender at seedling stage, identification of molecular markers to test genetic/clonal fidelity and incorporation of genes for genetic amelioration are the issues that need immediate consideration. Some of these issues have been addressed in the present paper. Keywords: Actinidia, conservation, genetic transformation, in vitro techniques, kiwifruit, molecular markers IPC Code: Int. C1.7 A 01 H 4/00, 5/00;C 12 N 15/9 Introduction successfully". Initially, only seven cultivars of A. The genus Actinidia is a perennial vine, bearing deliciosa var. deliciosa were imported from New berry fruits. It contains about 60 species, distributed Zealand and USA. However, the commercial potential in the temperate and subtropical regions of the Asian and suitability in the mid and foothills of Himachal continent'. Of these, A. deliciosa var. deliciosa and A. Pradesh further led to their large-scale propagation. chinensis are the only two species bearing edible During the last three decades, success in marketing of fruits. Further, A. deliciosa (A.Chev.) c.F. Liang et selected cultivars of A. deliciosa has also initiated A.R. Ferguson, commonly known as kiwifruit, is the breeding programmes aimed at developing novel only species cultivated all over the world. The first commercial cultivars from a large range of variation commercial cultivation of kiwifruit began in New available in this genus. Under such programmes, Zealand and, thereafter mostly during 1970s, it spread traditional breeding along. with molecular to other countries. Presently, New Zealand has the technologies is expected to result in genetically best-developed kiwifruit industry with 12,000 ha of improved new cultivars. Efforts have also been made land under its cultivation, out of total world in the field of its molecular biology, such as detection cultivation of 22,900 ha". The land under kiwifruit of molecular markers and genetic transformation, to plantations in New Zealand has further increased to ameliorate the genetic make up of this novel fruit 15,000 ha in 1992, producing almost 230,000 tons crop. However, dioecy and long juvenile period in fresh fruie. kiwifruit are the other equally important constraints of In India, kiwifruit was introduced in 1960s in parts breeding programmes. of the Himalayan region. Where it has been grown Taxonomy /Evolution The genus Actinidia, containing about 60 species, *Author for correspondence: Tel: 91-1792-52310; Fax: 91-1792-52242 can be further divided into varieties and forms. The E-mail: [email protected] two edible species, A. deliciosa and A. chinensis are 250 INDIAN J BIOTECHNOL, APRIL 2004 considered closely related. A. deliciosa is hexaploid Kumar and Sharma II described in detail the important and is probably derived from A. chinensis, which is features of rnicropropagation, i.e. the choice of diploid or tetraploid". Other species with economic explant, establishment of aseptic cultures, proli- potential are A. arguta, A. eriantha, A. latifolia, feration, rooting and hardening. Micropropagation by A. chrysantha, A. melandra, A. kolomikta and axillary bud stimulation has successfully been A. polygama. However, they are not commercial fruit employed for production of a large number of crops as yet. uniform kiwifruit plants from a single explant in a The origin of kiwifruit plant in New Zealand can short period of time. Successful shoot-tip culture of be traced back to two female and one male plants, mature plants has also been applied for vegetative introduced in 1904 from China in the form of seeds". propagation of staminate and pistillate plants". In 1920s, grafted plants of known sex were sold there and selections for high quality fruit were made". As a Organogenesis and Somatic Embryogenesis The regenerative potential of kiwifruit cells has in result, the well-known cultivars of today, viz. ~bbot', ~llison', 'Bruno', (Monty', ~ayward' and general been found to be fairly high irrespective of the tissue source. Leaf segments, stem pieces, petioles <Gracie', originated as seedling selections in New and root tissue have been reported to be amenable for Zealand. «JIayward' has now attained pistiIlate callus formation and subsequent plant regeneration. cultivar of commercial importance, for its better Cultivar 'Hayward' clones had higher proliferation shape, size, and long storability of fruits, not only in rates than those of the cultivar 'Tomuri'. Root New Zealand but also in the rest of world". Breeding cultures of these two cultivars have also shown high programmes for Actinidia, which currently has regenerative potential.". Endosperm tissue has also sizeable commercial industry, are being set-up in New been successfully cultured to produce triploid plants Zealand and Italy. (2n=3x=87)31. Another species, A. arguta also Propagation exhibited high morphogenic potential from various Propagation by seed can easily be carried out in explants such as internode, mature and immature kiwifruit but it results in a highly variable progeny leaves, petioles, immature seed and fruit, and in vitro and sex of the seedlings remains unidentifiable tiIl differentiated roots'". Internodal segments of A. 33 flowering. Therefore, vegetative propagation is kolomikta and petiolar sections of A. deliciosa cv. 34 considered superior to maintain uniformity and 'Hayward' showed high morphogenetic potential . genetic purity of the cultivars. Kiwifruit is generally Apart from these normal tissues as explant, plants propagated by grafting, budding, softwood and were also successfully regenerated from the callus hardwood cuttings, etc. However, large-scale planting derived from fruit galls of A. poiygama", The root materials are needed to meet the commercial require- tips of such regen eran ts showed 2n=58 as the ments. Hence, a simpler, quicker and economically chromosome number, which was the same as that of viable technique has to be needed for mass the mother plant. propagation. Several investigators have suggested Marino and Battistini, while comparing pH effects micropropagation as the best alternative for kiwifruit of the culture media, observed that though pH above propagation 7-11. Micropropagation and plant 5.7 increased the callus growth yet it inhibited the regeneration from callus was first tried in kiwifruit plant regeneratiorr'", The morphogenetic responses of I3 over a quarter century agoI2. Now, plants can be the explants also vary with the type, combination and regenerated from a variety of explants, such as apical concentration of growth regulators added to the basal meristem':', apical parts of shoots", axillary buds", medium. Canhoto and Cruz cultured pieces of young bark", cotyledons", endosperm'Y", internodes", leaf leaves from female and male plants of A. chinensis": segments 21,no des an d petio. Ies 22,stamens 23,and stem The explants were. cultured on MS medium with segments'r". Whereas, nodal segments or shoot tips auxin (NAA) and cytokinin (BAP). The buds arose are the most commonly used initial explants. Also, for from swellings on the cut ends of the leaf veins and 25 in 'vitro multiplication of kiwifruit, MS , Cheng's" differentiated into shoots. MS medium with NAA 27 and Gambog B5 media have so far been used. (0.1-0.5 mg/l) and BAP (2 mg/l) gave the best results. l2 Harada was the first to develop a protocol for The shoots rooted well in MS medium with 1 mg/l micropropagation of kiwifruit, which was later IBA and produced normal plants. Only minor improved by others28.29. Further, Standardi", and differences in the morphogenetic potential among SHARMA & SHIRKOT: GENETIC AMELIORATION OF KIWIFRUIT PLANT 251 explants of different origins were observed. Similar significantly greater than those of hexaploid observations, that nature of the explant did not affect somaclones. Using callus cultures of cv. 'Hayward', plant regeneration from callus, were also reported in somaclones were regenerated with increased tolerance cv. 'Hayward=", to high pH as compared to the control'". These clones Somatic embryogenesis can also be used for rapid also retained the pH tolerance under field conditions. clonal propagation of homogeneous transgenic plants. Further, testing of these materials under field However, inconsistency in embryo induction in conditions resulted
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