Improved in vitro propagation of sweetpotato [Ipomoea batatas (L.) Lam.] – Confirmed with a wide range of genotypes (original) (raw)
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Plant Cell, Tissue and Organ Culture (PCTOC)
In vitro propagation of Plant Genetic Resources is a basic step for routine genebank and biotechnology research activities. Accelerating growth and rooting of in vitro plants contributes to an improvement in process efficiency and plant quality. In the present study the effect of supplemental thiamine and explant size on biometric variables, ion content in plant sap, chlorophyll content in leaves and moisture content in plants were assessed in a replicated trial on a group of seven in vitro sweetpotato accessions and validated in a set of other 45 accessions. It was shown that adding 0.1 mg L −1 of thiamine to modified Murashige and Skoog culture medium significantly increased plant height, root length, and number of nodes of in vitro sweetpotato shoot culture plants. No significant differences were observed for N03−, K+, Na+ and Ca++—ion content in plant sap, nor in leaf area, chlorophyll, or moisture content between plants grown with or without thiamine. Uninodal stem segments sho...
Chemical and environmental growth regulation of sweetpotato (Ipomoea batatas (L.) Lam.) in vitro
Plant cell, tissue and organ culture, 1991
The need for conservation of biotic diversity is well recognized. However, improved techniques for the efficient, cost effective-preservation of plant germplasm are needed. The conservation and distribution of plant germplasm in vitro is gaining acceptance. However, increased usage is dependent upon the ability of curators to minimize culture maintenance requirements. This report examines the effect of various levels of sucrose, photoperiod, temperature, sorbitol and mannitol on minimal growth storage of Ipomoea batatas (L.) Lam. Growth was reduced 50% with a temperature reduction of from 21.1 to 15.6°C. Sucrose concentrations of 15 and 20 g 1-1 resulted in reduced plant stature with few adverse effects on plantlet viability or morphology. Reduction of photoperiod from 16 to 4 h produced smaller, slightly chlorotic, but otherwise normal plants. The addition of sorbitol or mannitol to culture media generally produced undesirable effects on gross plant morphology and loss of apical dominance. Genotype x growth retarding treatment interactions were observed for all variables examined.
In Vitro propagation of sweet potato (Ipomoea batatas (L.) Lam) through apical meristem culture
Sweet potato (Ipomoea batatas L.) is an important warm season crop that grows best in long, hot growing seasons, cultivated in over all developing countries as a valuable source of human food, animal feed and industrial raw material. Its yield is highly decreased due to disease in Eastern African Countries. Production of virus free sweet potato is of great potential to increase yield and improve the crop. In the present study, different treatments, each with a combination of (BAP and Kn) in a ratio 0.25, 0.5, 0.75, 1.0 and 0.25, 0.5, 0.5, 0.75 and 0.0 mg/l (control) and auxin (IBA and NAA) in a ratio 0, 0.5, 0.5, 1.0, 1.0mg/l and 0, 0.25, 0.5, 0.5 and 1.0mg/l in combination respectively were used for three varieties (Beletech, Awassa-83 and Belela) to optimize the concentrations of BAP, Kn, IBA and NAA in MS basal media for meristem culture. The experiment was laid out in CRD with three replications in factorial arrangement. Among the growth regulator combinations tested 0.5mg/l BAP with 0.5mg/l Kn followed by 0.75mg/l BAP with 0.5mg/l Kn were best for all shoot induction and growth parameters for the three varieties. The minimum days to root induction was recorded for Beletech (3.167days) shoots cultured on media supplemented with 0.5mg/l IBA with 0.5mg/l NAA and Awassa-83 (3.83days) shoots cultured on media supplemented with 1.0mg/l IBA and 0.5mg/l NAA. Whereas, the variety Belela shoots have induced in (3.83 days) on media with 1.0mg/l IBA with 1.0mg/l NAA. Maximum number of roots per shoot were recorded on MS media supplemented with a combination of 1.0mg/l IBA with 0.5mg/l NAA (11.7) followed by (9.3) on media with 0.75mg/l IBA with 0.5mg/l NAA respectively. The maximum root length was observed for Beletech (3.4cm) followed by Awassa-83 (3.43cm) cultured on the media with a combination of IBA 0.75mg/l with NAA 0.5mg/l. The results indicated that better response was obtained for root related parameters of all variety shoots transferred for rooting on media supplemented with 0.5mg/l IBA with 0.5mg/l NAA and above up to 1.0mg/l IBA with 0.5mg/l NAA. During acclimatization, 90%, for Beletech and 80% for Awassa-83 and Belela varieties survival was obtained.
Journal of Tissue Science & Engineering, 2016
Sweetpotato is an important food crop in the world as well as in Kenya. Various fungal and viral diseases are major constraints in its production and are currently threatening the sweetpotato production in sub-Saharan Africa. Genetic engineering offers significant potential for the crop's genetic improvement. However, this is limited by the low efficiency and strong genotype dependency in tissue culture. This study aimed to establish an efficient somatic embryogenesis and plant regeneration system using shoot apical meristem explants of sweetpotato. Three sweetpotato cultivars that are widely grown in Kenya; KSP36, Kemb36 and Mweu mutheke along with an exotic model cultivar Jewel were evaluated. The maximum somatic embryogenic induction, at 96.72%, was obtained from explants cultured on Linsmaier and Skoog salts and vitamins medium supplemented with 0.5 mg/l dichlorophenoxyacetic acid and 0.2 mg/l zeatin riboside. The highest number of shoot induction (33) was observed after transfer of embryonic callus to embryo maturation medium supplemented with 2 mg/l abscisic acid. Significant differences were observed between cultivars for somatic embryogenesis and plant regeneration. Jewel showed the best response, while Mweu mutheke was the least responsive under the culture conditions tested in this study. Regenerated plants were successfully rooted and grown to maturity after hardening in soil in the greenhouse. Such a robust, successful and efficient system possesses the potential to become an important tool for crop improvement and functional studies of genes in sweetpotato.
Thidiazuron-induced adventitious shoot regeneration of sweetpotato ( Ipomoea batatas
in Vitro Cellular & Developmental Biology-plant, 1995
Adventitious shoots of sweetpotato (Ipomoea batatas L. Lam.) were produced in vitro using a two-stage culture method. Petiole explants were incubated on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (0.2 mg·liter−1) for 3 d, and transferred to MS medium with thidiazuron (0 to 0.4 mg·liter−1). Shoot regeneration was observed in most explants (78.2%) of genotype PI 318846-3 within 28 days when cultured on thidiazuron at 0.2 mg·liter−1. Histological studies of cultured petiole explants showed meristematic activity within cells of vascular bundles and throughout the ground tissue. Explants isolated from apical leaves exhibited higher shoot regeneration frequency than those isolated from the basal portion of the shoot. Leaf lamina explants exhibited lower frequency of regeneration than petiole explants. In contrast to thidiazuron, the use of zeatin riboside, and kinetin resulted in a lower frequency of shoot regeneration although more sweetpotato genotypes could be regenerated using either of these two cytokinins. The sweetpotato plants regenerated using thidiazuron grew vigorously and rooted easily when transferred to the greenhouse.
Scientia Horticulturae, 1995
Leaf explants of sweetpotato (1pomoea Bahamas L. (Lam.) ) were cultured on Murashige and Skoog medium (MS) with varying levels (0.2-l mg l-') of 2,4-dichlorophenoxyacetic acid (2,4-D; stage I) and transferred to a medium with zeatin riboside (0.2-0.4 mg l-i; stage II). The highest frequency (over 80%) shoot regeneration occurred in the genotype PI 318846-3 cultured on MS + 2,4-D (0.2 mg 1-i) for 3 days and then transferred to MS +zeatin riboside (0.2 mg 1-i). Addition of zeatin riboside in the first medium reduced the frequency of shoot regeneration. Of the 27 genotypes, 19 exhibited high frequency shoot regeneration, and eight were completely recalcitrant. The developmental stage of the leaf was critical, as young leaves from the apical portions of the stem were the most regenerative. Regenerated shoots rooted readily and the plants transferred to soil in the greenhouse appeared normal. Abbreviations: 2,4-D = 2,4-dichlorophenoxyacetic acid; ABA = Abscisic acid 0304-4238/95/$09.50 0 1995 Elsevier Science B.V. All rights reserved SSDIO304-4238(95)00767-9
Induction of somatic embryogenesis in recalcitrant sweetpotato (Ipomoea batatas L.) cultivars
AFRICAN JOURNAL OF BIOTECHNOLOGY
Genetic transformation is considered as one of the most promising options for improvement of crop traits. Current transformation methods for sweetpotato depend on plant regeneration through organogenesis or somatic embryogenesis. Somatic embryogenesis and plant regeneration at a high frequency has been restricted to a few sweetpotato varieties. Three auxins namely: 2,4dichlorophenoxyacetic acid (2,4-D), 4-fluoroamphetamine (4-FA) and 4,5-trichlorophenoxyacetic acid (2,4,5-T) were investigated in this study for enhancing somatic embryogenesis from various plant organs of recalcitrant African sweetpotato cultivars. 2,4-D was found to be the best (p ≤ 0.05) for induction of embryogenic callus. Cultivar Bwanjule had the highest (20.2%) embryogenic callus frequency among the five African cultivars tested. The highest number of plants in this study was regenerated from the non-African cultivar variety Jonathan on media supplemented with 0.2 mg Zeatin. The emergence of roots from callus of recalcitrant Ugandan cultivars and the comparable high embryogenic responses in this work demonstrate the potential for regenerating plants from African cultivars that have not been regenerated before. The regeneration of roots in this work could be useful for the initiation of root cultures. The most important application of this work is in genetic transformation of sweet potato, particularly for improvement of resistance to weevils.
In vitro propagation of sweet potato (Ipomoea batatas (L.) Lam.) cultivars
Agricultural Science Digest - A Research Journal, 2018
The study is aimed at establishing a simple protocol for in vitro regeneration of sweet potato with a view to providing planting materials to farmers as well as basis for genetic improvement. Axillary buds were excised and cultured on Murashige and Skoog (MS) basal salts supplemented with 6-benzyl aminopurine (BAP), gibberellic acid (GA3) and naphthalene acetic acid (NAA) singly or in combination. The shoot height and number of leaves differed significantly among the cultivars. The result also indicated significant difference (p less than 0.01) among the cultivars with King J recording the highest mean values. Significant differences (p less than 0.05) was also recorded in the media combination with respect to organogenesis and number of shoots obtained. The results of hardening further revealed 33.33% success in the explants transferred directly to the field, as well as for the plantlets that were gradually weaned in a mixture of 3:1 sand and biochar.
Plant Cell Tissue and Organ Culture, 2009
A storage root-bearing somatic hybrid was produced for the first time by protoplast fusion between sweetpotato (Ipomoea batatas (L.) Lam.) cv. Kokei No. 14 and its wild relative I. triloba L. Protoplasts isolated from embryogenic suspension cultures of Kokei No. 14 were fused with petiole protoplasts of I. triloba L. using polyethylene glycol-mediated protocol. Fusion products were cultured in a modified Murashige and Skoog medium containing 0.05 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg l−1 kinetin. A total of 176 plants were obtained from 42 out of 134 calluses derived from fused protoplasts, and 91 of these plants were confirmed to be somatic hybrids through peroxidase isozyme, random amplified polymorphic DNA, amplified fragment length polymorphism, and cytological analyses. Upon transfer into soil and grown in the greenhouse and then to the field, 100% survival was observed. A single plant, designated KT1, was found to produce storage roots. Genomic in situ hybridization analysis confirmed presence of chromosomes from both parents and recombinant chromosomes in KT1. Drought tolerance, dry matter content, soluble sugar content, and fertility of this somatic hybrid were evaluated for potential use in sweetpotato breeding.