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Journal ArticleDOI

Morphogenetic investigation on pea under in vitro conditions

01 Oct 1985-Bulletin of the Torrey Botanical Club (Torrey Botanical Club)-Vol. 112, Iss: 4, pp 363-367
About: This article is published in Bulletin of the Torrey Botanical Club.The article was published on 1985-10-01. It has received 8 citations till now.
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Journal ArticleDOI
TL;DR: The transmission of the introduced gene into the progeny of the regenerated transgenic plants was studied over two generations, and stable transmission was shown to take place.
Abstract: An analysis of the progeny of primary transgenic pea plants in terms of transmission of the transferred DNA, fertility and morphology is presented. A transformation system developed for pea that allows the regeneration of fertile transgenic pea plants from calli selected for antibiotic resistance was used. Expiants from axenic shoot cultures were co-cultivated with a nononcogenic Agrobacterium tumefaciens strain carrying a gene encoding hygromycin phosphotransferase as selectable marker, and transformed callus could be selected on callus-inducing media containing 15 mg/l hygromycin. After several passages on regeneration medium, shoot organogenesis could be reproducibly induced on the hygromycin resistant calli, and the regenerated shoots could subsequently be rooted and transferred to the greenhouse, where they proceeded to flower and set seed. The transmission of the introduced gene into the progeny of the regenerated transgenic plants was studied over two generations, and stable transmission was shown to take place. The transgenic nature of the calli and regenerated plants and their progeny was confirmed by DNA and RNA analysis. The DNA and ploidy levels of the progeny plants and primary regenerants were studied by chromosome analysis, and the offspring of the primary transformants were evaluated morphologically.

51 citations

Journal ArticleDOI
TL;DR: A sequential indole-3-acetic acid-zeatin treatment was applied to Pisum sativum hypocotyl explants, resulting in shoot formation from 50% of theExplants, which suggests the shoots to be of de novo origin, which would make the system suitable for transformation experiments.
Abstract: A sequential indole-3-acetic acid (IAA)-zeatin treatment was applied to Pisum sativum hypocotyl explants, resulting in shoot formation from 50% of the explants. Shoots were easily rooted and transplantable plants could be obtained in 3 months. The method has been applicable to the 5 cultivars tested. Histological examination of explants suggests the shoots to be of de novo origin, which would make the system suitable for transformation experiments.

16 citations

DOI
01 Jan 2010
TL;DR: The result showed that combination of BAP and NAA was able to induce callogenesis from hipocotyl of alfalfa and the optimal callus was obtained in combination of NAA 2 ppm and BAP 0 ppm.
Abstract: Alfalfa (Medicago sativa L.) is useful plant for treatment some diseases such as: cancer, diabetes, lupus, and hepatitis. Propagation of this plant in Indonesia face a problem which has no embryo. One method to propagate this plant is by tissue culture or micropropagation. Callus induction is first step in micropropagation to produce callus which will be regenerated to become planlet. The aims of this research are to induce callogenesis from hipocotyl of alfalfa with addition Benzyl Amino Purine (BAP) and α Naphtalene Acetic Acid (NAA), and to determine the proper combination of BAP and NAA to produce the optimal callus. The experiment has been conducted by using 12 combination of BAP and NAA with Completely Randomized Design (CRD) in 4x3 factorial pattern by 5 replicates. Data were analyzed by ANOVA 95% Degrees of Freedom (DF). If there was significance result, it was followed by DMRT analyzed at 95 % DF. The result showed that combination of BAP and NAA was able to induce callogenesis from hipocotyl of alfalfa. The optimal callus was obtained in combination of BAP 0 ppm and NAA 2 ppm.

12 citations

Journal ArticleDOI
TL;DR: Variations of morphogenetic behaviour observed in meristem cultures of the investigated pea cultivars manifested a rather strict genetic determination of their regeneration abilities, especially in respect of flowering and rooting of the regenerated shoots.

9 citations

Journal ArticleDOI
TL;DR: These experiments suggest that mutations affecting developmental characters may be expressed in tissue culture and may cause increased adaptation in mutant cells to the in vitro conditions, which may lead to a higher proportion of quantitative mutations among the regenerated plants.
Abstract: A study of regenerants obtained from long-term callus cultures of different pea (Pisum sativum L.) genotypes revealed specificity of in vitro mutagenesis. The specificity was displayed in the preferential generation of the somaclonal variations affecting quantitative and developmental characters. About 60% of regenerated lines obtained from the cultivar Ranny Zeleny carried mutations in the Lf and Sn loci, which control initiation of flowering. An in vitro study of isogenic lines differing at the Lf and Sn loci illustrates the growth advantages of mutant genotypes in tissue culture. These experiments suggest that mutations affecting developmental characters (e.g. mutations in loci which control flowering behaviour) may be expressed in tissue culture and may cause increased adaptation in mutant cells to the in vitro conditions. Rapid propagation of mutant cells during in vitro culture may lead to a higher proportion of quantitative mutations among the regenerated plants.

7 citations