Plant Cell Culture in Crop Improvement

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In Vitro Tissue Culture S tudies in Sunflower (Helianthus spp.)

References Publications referenced by this paper. In vitro fertilization and embryo rescue. Berlin Springer Verlag.

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Yeung , E. C , Thorpe , TA. Ex situ conservation of plant germplasm using biotechnology Victor M. Villalobos , Florent Engelmann. In vitro production of haploid plants Atanas Atanassov , N. Zagorska , P. Boyadjiev , Dimitar Djilianov. Recent advances in plant cell cultures in bioreactors. Zhong , J. Somatic embryogenesis in herbaceous dicots. The explant is then further reduced in size 0.

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  7. Meristem cultures have the disadvantage that they may have a higher mortality rate and an initial slower growth. The explant is placed directly on a multiplication-inducing culture medium. For banana micropropagation, MS-based media [8] are widely adopted. Banana tissue cultures often suffer from excessive blackening caused by oxidation of polyphenolic compounds released from wounded tissues. These undesirable exudates form a barrier round the tissue, preventing nutrient uptake and hindering growth.

    Therefore, during the first weeks, fresh shoot-tips are transferred to new medium every weeks. Alternatively, freshly initiated cultures can be kept in complete darkness for one week. Usually two types of growth regulators, a cytokinin and an auxin, are added to the banana growth medium.

    Plant Tissue Culture and Genetic Transformation for Crop Improvement

    Their concentration and ratio determines the growth and morphogenesis of the banana tissue. We routinely add 2. In most banana micropropagation systems, semi-solid media are used. Liquid media are superior for shoot multiplication [10], but for maximum plant production and survival ex vitro , one culture cycle on semi-solid medium is also needed. The formation of multiple shoots and buds is promoted by supplementing the medium with relatively high concentrations of cytokinins.

    In banana, BA is the preferred cytokinin and is usually added in a concentration of 0.

    From Lab to Field – Improving a Major Staple Crop for Food Security

    For the multiplication of propagules, we use the same medium as for the initiation of shoot cultures p5 medium containing 2. If the production of highly proliferating meristem cultures is required Section 3. Higher concentrations of the cytokinin BA tend to have an adverse effect on the multiplication rate and morphology of the culture and should therefore be avoided.

    The rate of multiplication depends both on the cytokinin concentration and the genotype. In general, shoot tips of cultivars having only A genomes produce new shoots, whereas cultivars having one or two B genomes produce a cluster of many shoots and buds at each subculture cycle.

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    An Introduction to Plant Tissue Culture: Advances and Perspectives

    Approximately weeks after culture initiation, depending on the initial explant size, new axillary and adventitious shoots may arise directly from the shoot-tip explant. Clusters can be separated, trimmed and repeatedly subcultured at week intervals. Individual shoot or shoot clumps are transferred to a nutrient medium which does not promote further shoot proliferation but stimulates root formation. The cytokinin in the regeneration medium is greatly reduced or even completely omitted. Within 2 weeks, shoot tips develop into unrooted shoots.

    Crop improvement through tissue culture

    We use the same auxin concentration as in the proliferation medium 0. For some genotypes Musa spp. ABB and BB group that produce compact proliferating masses of buds, activated charcoal 0. Shoot or meristem tip cultures are suitable not only for the large-scale production of uniform and vigorously growing propagules for field establishment, but also for germplasm conservation.

    For banana species which are predominately propagated vegetatively, in vitro techniques are thus complementary to field conservation. Hence there is the risk of losing material through microbial contamination or through human error mislabelling of culture vessels. Nevertheless, conservation of germplasm under short-term storage conditions is most useful, for instance, to breeders or researchers who wish to maintain a small working collection of breeding material during evaluation, testing, selection and hybridisation, or to maintain products of in vitro manipulation.

    Slow growth conservation has the benefit of reducing the number of subcultures, thus making significant savings in labour input.

    Application of tissue culture in crop improvement

    Germplasm still remains readily available for regeneration, multiplication and distribution. Minimal growth is clearly useful for conservation of genotypes, but is constrained by the risk of genetic changes somaclonal variation resulting in the loss of distinct genotypes. The most common and widely applied growth-retarding factor in banana is low temperature. Subculture intervals are extended to about 12 months, although large differences in storage potential have been observed among the different genotypes: some genotypes keep for about 20 months whereas others require subculturing every 2 months [12].

    Alternatively, the use of osmotica in banana tissue cultures has been investigated [13]. Contamination in tissue cultures may be caused by endogenous bacteria that escape initial disinfection or by micro-organisms introduced during tissue-culture manipulations. Both types of contaminants may survive in the plant material for several subculture cycles and over extended periods of time without expressing symptoms in the tissue or visible signs in the medium.

    It is therefore important that control measures are taken at every tissue-culture step. At the INIBAP Transit Centre, plant material is tested for endophytic bacteria on a broad spectrum bacteriological medium at tissue culture initiation and during annual subculturing. In mass propagation systems, positive stock materials should immediately be destroyed. However, germplasm in collections or cultures that are not readily replaceable with fresh material e. Some antibiotics are successful in controlling bacterial contaminants in banana tissue cultures. However, culturing small meristem tips 1 mm isolated from contaminated in vitro plants or from greenhouse plants obtained from contaminated in vitro plants was found to eliminate any bacterial contaminant [15].

    Shoot-tip culture preserves genetic stability much better than callus or cell suspension cultures, yet somaclonal variation appears to be widespread among plants regenerated from banana shoot-tip cultures.

    In Vitro Production of Triploids

    The phenomenon of somaclonal variation in the plantain subgroup Musa spp. AAB group was extensively studied [17]. It revealed that the incidence of somaclonal variation is strongly influenced by the genetic stability of each cultivar, and that its frequency is amplified by culture-induced factors.

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    There is no evidence that growth regulators routinely used in tissue culture directly affect the rate of variation, but it has been found that the rate of somaclonal variation is positively related to the generation number. For 'Williams' AAA in vitro plants obtained after one and five subculture cycles, dwarfism and leaf-off types counted for 3.

    It is therefore recommended that the number of subculture cycles should be limited to ten [19] or that the number of plants produced from a primary explant should be limited to no more than [20]. In bananas and plantains, there are four main procedures for the development of embryogenic cell suspensions. They differ mainly by the explant in which embryogenesis is induced: zygotic embryos [21, 22], rhizome slices and leaf sheaths [23], immature fe male flowers [], and proliferating meristem cultures [27, 28].

    Each method has its own limitations, rendering the establishment of Musa embryogenic cell suspensions still far from routine [29]. Since most edible banana cultivars rarely set seeds, zygotic embryos are of limited value as starting material. Reports on the efficiency of embryogenesis induction in highly differentiated rhizome and leaf tissue are scarce.