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Embryo Rescue in Wide Crosses in Arachis. 1. Culture of Ovules in Peg Tips of Arachis hypogaea
Abstract
Interspecific hybridization in Arachis is restricted by early embryo abortion for many cross-combinations. Rescue of young embryos in vitro within a week after fertilization is necessary to recover these embryos before they abort. Peg tips, with the ovule and embryo tissues, of A. hypogaea L. cv. 'NC 6', were cultured to compare ovule growth, callus production and peg elongation. Tissues were collected 1, 2, 3 and 4 d after self-pollination, after which peg meristems were removed from half the pegs and cultured on five media combinations. One-day-old pegs had significantly (P = 0.01) more ovule growth than older tissues. Presence of the meristem had a greater inhibition to ovule growth for 2- to 4-d pegs than for 1-d-old pegs. Significantly more callus was produced on 4-d pegs than younger tissues, and kinetin had the greatest stimulatory effect on callus. Elongation of pegs with the meristem attached was observed most often in media with high sucrose levels. The observations indicate that very young ovules can be grown in vitro, and techniques may be applicable to rescue of young embryonic tissues of Arachis.
... Their results indicated that several selfed and hybrid embryos grew to the globular stage after 21 d in culture. Moss et al. (13), Pattee et al. (19), and Rau et al. (20) later found similar results with 1-to 4-d-old selfed peg tips of A. hypogaea. Ziv and Sagar (23) c u l t u r e d 4 to 5-cm pegs ofA. ...
... Embryo development from the globular to the heartshaped stage is a critical step for which in vitro techniques need to be developed (13,19,20). In th' IS research, large embrvos and mature seeds were recovered " from proembryos through a one-step in vitro procedure. ...
... and effects of plant growth regulators were studiLd. Previous attempts to culture peanut pegs have been primarily with short peg tips without meristems (12,13,19,20). Although the removal of meristems from Dew can eliminate inhibitorv I " effects of elongation on embryo development, the c; t surface is so close to the ovary that in vitro growth of ovules and embryos is probably influenced by wounding effects. ...
In vitro culture of embryos in Arachis is necessary to recover interspecific hybrids which otherwise abort soon after fertilization. The objective of this research was to develop in vitro techniques to promote proembryo de-velovment so that ~lants can be recovered. Aerial Deg. 1 1 7 tips consisting of embryos, ovules, and peg meristem of Arachis hypogaea L. cv. NC 6, were collected 7,10, and 14 d after self-pollination. Peg tips were cultured in the dark on combined MS and B5 media with NAA, GA, and 6-BAP for 90 d. The effects ofplant growth regulators on in oitro reproductive traits, including peg elongation, callus and root production, pod formation, ovule and embryo development were variable. Results indicated that 10-d-old peg tips, which contained eight-celled proembryos, had more embryo development and pod formation than 7-and 14-d-old peg tips. Medium with 4 mg L-' NAA and 0.5 mg L-I 6-BAP suppressed in vitro development of pods, ovules and embryos and induced large amounts of callus. Media with lower concentrations of NAA, GA,, and 6-BAP caused development of more and larger pods and ovules. The development of young embryos from proembryos was observed and mature seeds were obtained by an in vitro one-step process. Peanut plants were obtained both from in uitro-recovered embryos and from mature seeds.
... Embryo rescue from wide crosses in Arachis Moss et al. (1988) Moricandia arvensis X Brassica ...
... In case of seed sterility, the plantlets are treated with colchicine for production of amphiploids through chromosome doubling. It has been a practical approach (Sharma et al. 1996) to obtain interspecific and intergeneric hybrids in Arachis (Moss et al. 1988), banana (Dayarani et al. 2014), Brassica (Takahata and Takeda 1990;Momotaz et al. 1998;Gupta et al. 2010;Ding et al. 2011;Mithila and Hall 2012;Chamola et al. 2013;Sun et al. 2014), chickpea (Clarke et al. 2011), citrus (Aleza et al. 2010;Liu et al. 2010a;Kurt and Ulger 2014), cucumber (Plapung et al. 2014), Cucurbita (Rakha et al. 2012a, b), grapevine (Wang et al. 2010;Donici and Tardea 2012;Niu et al. 2012;de Menezes et al. 2014), Helianthus (Chandler and Beard 1983;Sauca 2010;Sauca and Lazar 2011), Lens (Cohen et al. 1984;Suvorova 2014), Lilium (Van Tuyl et al. 1991), Populus (Calagari et al. 2004;Payamnour et al. 2013), sesamum (Rajeswari et al. 2010), sorghum (Rizal et al. 2014), Vigna (Gosal and Bajaj 1983) and wheat (Kaur et al. 2002;Sehgal et al. 2011). Interspecific and intervarietal hybrids have been generated in seedless citrus, seedless grape, mango and papaya using embryo rescue method. ...
Plant cell and tissue culture involves the growing of cells, tissues and organs on synthetic medium under closely controlled and aseptic conditions. Plant cell and tissue culture methods offer a rich scope for the creation, conservation and utilization of genetic variability for the improvement of field, horticultural and forest plant species. Micropropagation of selected plant species is one of the best and most successful examples of the commercial application of tissue culture technology. Micropropagation ensures true-to-type, rapid and large-scale multiplication. Now scores of multimillion-dollar industries around the world propagate a variety of plant species through micropropagation. Tissue culture technology offers environmental-friendly industries to flourish. It is likely that automation of multiplication systems will be commercially feasible within the next few years for several species including potato microtubers, lily bulblets and gladiolus corms. Meristem culturing and in vitro grafting help in developing disease-free plants. Improvement of somatic embryogenesis, coupled with embryo desiccation and encapsulation technology, may lead to the utilization of ‘artificial seeds’ for mass cloning of plants. Further induction of somatic embryogenesis in plants helps in cloning and transformation. Somaclonal variation is a potent emerging aspect for broadening the genetic base and thus obtaining incremental improvement in the commercial cultivars, more particularly, in the vegetatively propagated plant species. Using the technique of in vitro selection, many million cells/protoplasts can be screened against various biotic and abiotic stress factors in a single Petri dish which is more efficient as compared to the screening of similar number of plants in the field which requires more time and space as well. Production of haploids through bulbosum, anther/pollen culture and embryo rescue from wide hybrids has been exploited for the production of haploids/doubled haploids for early release of varieties. These methods ensure true-breeding (doubled haploids) plants in less than 1 year, which are otherwise obtained after seven to eight generations through conventional methods. Since the possibility of producing useful secondary products in plant cell cultures was first recognized in the 1970s, considerable progress has been made, and a number of plant species have been found to produce secondary products such as shikonin, diosgenin, caffeine, glutathione and anthraquinone. Embryo culture is the practical approach to obtain interspecific and intergeneric hybrids among otherwise difficult to cross parents. It has been successfully used to transfer desirable genes from wild relatives into cultivated varieties of several field and vegetable crops. Somatic cell hybridization helps in combining characteristics even from otherwise sexually incompatible species and to obtain cybrids and organelle recombination not possible through conventional methods. In vitro freeze storage and cryopreservation are very important techniques for germplasm conservation especially of the vegetatively propagated crops. Plants have been successfully regenerated from tissues cryopreserved at –196 °C in liquid N2 for several months to years in several crops. During the past 25 years, the combined use of recombinant DNA technology, gene transfer methods and cell and tissue culture techniques has led to the efficient transformation and production of transgenics in a wide variety of crop plants. In fact, transgenesis has emerged as an additional tool to carry out single-gene breeding or transgenic breeding of crops.
... Previous work has resulted in multicellular globular embryos derived from one-to two-celled proembryos of selfed A. hypogaea L. (8,13,14). Ziv and Sagar (23) achieved in vitro growth of A. hypogaea ovules and obtained viable young seedlings by a two-step process. Mature seeds were obtained from several-celled proembryos of A. hypogaea and A. duranensis Krapov. ...
Research on in vitro embryo culture in Arachis has the objective ofrescuinginterspecific hybridembryos which abort before they reach maturity. This study explored effects ofthe three exogenous plant growth regulators 1- naphthaleneacetic acid (NAA), gibberellic acid (GA,), and 6-benzylaminopurine (6-BAP); sucrose; and me- dium pH on in vitro fruit and embryo development ofA. hypogaea L. by culturing 10-d-old peg tips. Results indicated that medium containing 0.5 to 1.0 mg L NAA was optimal for in vitro pod formation and embryo development. GA, did not have a significant influence and 6-BAP had negative effects on both in vitro fruit and embryo development. High concentrations of 6-BAP and NAA induced callus which inhibited ovary enlarge- ment and embryo development. Sixty g L-' sucrose was the best concentration for ovary enlargement and em- bryo development. Acidic medium was needed for in vitro reproductive development with pH 4.5-6.5 the most favorable. A pod formation frequency of 81%, a seed production rate of 90% (from pods recovered in
In vitro culture of Arachis peg tips have been used to recue aborting embryo of interspecific hybrid, but increased efficiency is still needed. The objective of this research was to investigate whether glutamine can be supplemented to enhance podding and embryo development of Arachis hypogaea L. Fourteen-day-old, peg tips, with embryos and ovules, of cv. TN11 were cultured on the MS media combined with various glutamine levels. The results indicated that the basal MS medium was not sufficient to induce larger pods and mature seeds. Media containing glutamine could significantly enhance the percentage of rooting, ovary swelling and pod formation. Fifty mg/l glutamine was the most favorable concentration for pod formation and embryo development, but the induced response was different among cultivars. In media with added glutamine, embryos in the heart-shaped to late cotyledonary stage was observed and mature seeds was obtained. These observations illustrate that the complete developmental processes from ovaries to fruits and from proembryos to mature seeds in peanut can be achieved through a one-step in vitro procedure, and techniques may be applicable to rescue of interspecific hybrids.
Cross-incompatibility between cultivated peanuts and their wild relatives outside the section Arachis has impeded the utilization of many species possessing high resistances or good qualities. Despite the great efforts made to culture immature ovules or embryos, few hybrid offspring have been obtained. In this study, gynophores from Arachis hypogaea L. pollinated with A. glabrata Benth. were cultured and F1 hybrids seeds were harvested, and F2 and F3 generations produced. The characters of F2 generation exhibited a wide range of segregation. Leaf peroxidase isozyme PAGE analysis revealed that the hybrids were quite different from their parents in relation to band number, width and isozyme activity. The zymograms of the hybrids and their parents were partially alike. This verified the authenticity of the hybrids.
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