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A kind of method for inducing regenerated seedlings of paprika explants

A technology for explants and regenerated seedlings, applied in the field of plant tissue culture, can solve the problems of low differentiation rate of adventitious buds, weak root regeneration ability, etc., and achieve the effects of low pollution rate and stable genetic traits

Active Publication Date: 2022-05-27
HAINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In view of this, the present invention provides a kind of method for inducing regenerated shoots of Capsicum pilosa explants, solves the problem that only a small amount of callus is produced and the differentiation rate of adventitious buds is very low and For the problem of weak root regeneration ability, the in vitro regeneration of velvet capsicum was realized for the first time, including the following steps:

Method used

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  • A kind of method for inducing regenerated seedlings of paprika explants
  • A kind of method for inducing regenerated seedlings of paprika explants
  • A kind of method for inducing regenerated seedlings of paprika explants

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Embodiment 1

[0019] Embodiment 1: a method for inducing regenerated seedlings from explants of pubescent pepper, comprising the following steps:

[0020] (1) Obtaining sterile seedlings: Take the seeds of Capsicum pubescens, a capsicum cultivar, and place them in a 50 mL centrifuge tube, add ultrapure water, and place them at 4°C for 24 hours. The soaking time is too short and the germination time is slow. If soaking time is too long, the pepper seeds will rot and the germination rate will decrease. Then add 75% ethanol to shake for 1min, rinse with sterile water twice, shake with 10-15% sodium hypochlorite solution for 20min, rinse with sterile water for 5 times, Finally, the sterilized seeds are placed on sterile filter paper to absorb water, and then inoculated into MS medium for cultivation to obtain sterile seedlings;

[0021] (2) Induction of adventitious buds: step (1) normal growth of sterile seedlings for 20d, cut 0.5-1cm stalked cotyledons as explants, inoculated into induction m...

Embodiment 2

[0025] Embodiment 2: the influence of different explants on the induction and differentiation of adventitious buds, wherein in step 2, respectively select cotyledon, hypocotyl and stalk cotyledon as explants, and the cotyledons are used as explants as can be seen from the results in Table 1. The browning is more serious, and a small amount of callus can be produced, and the induction rate is 26%. The hypocotyl also has the problem of browning and can produce a large number of calli, but the induction rate of adventitious buds is low, and the highest differentiation rate is nearly 3%. . As explants, the petiolate cotyledons can swell near the petiole end and produce adventitious buds, and the induced differentiation rate is over 46%.

[0026] Table 1 Effects of different explants on shoot differentiation

[0027]

Embodiment 3

[0028] Embodiment 3: the influence of different seedling ages on the induction and differentiation of adventitious buds, wherein in step 2, selecting seedlings of 15d, 25d and 30d aseptic seedlings of pubescent peppers to carry out excision tissue to induce and cultivate regenerated seedlings, the results are as shown in Table 2, When the sterile seedlings of 20-25 days old were selected, the induction rate of adventitious buds was high, reaching 48%, indicating that too large or too small seedlings of sterile seedlings were not conducive to the formation of callus and adventitious buds.

[0029] Table 2 Effects of different seedling ages on bud differentiation

[0030]

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Abstract

The invention discloses a method for inducing regenerated seedlings of capsicum pilosa explants, which comprises the steps of firstly obtaining aseptic seedlings, then cutting cotyledons with stalks as explants, inoculating them on induction medium to induce adventitious buds, and then cutting them sequentially Transfer to elongation medium and rooting medium, where MS induction medium is MS+6‑BA 5.0mg / L+IAA 0.5mg / L+AgNO 3 3.0mg / L; elongation medium is MS+6‑BA 2.5mg / L+IAA 0.8mg / L+GA 3 3.0mg / L+AgNO 3 3.0mg / L; the rooting medium is 1 / 2MS+NAA 0.2mg / L. After the rooted seedlings reach a height of 8-10cm, remove the bottle cap and place them under natural light at room temperature for hardening and transplanting. As explants, the in vitro regenerated seedlings with higher germination induction rate and rooting rate were obtained for the first time by applying the developed induction medium, elongation medium and rooting medium. The in vitro regeneration system constructed by the method of the present invention is The gene function verification and molecular breeding of capsicum paprika laid the research foundation.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for inducing and regenerating seedlings from explants of pubescent peppers. Background technique [0002] Tissue culture is an important part of agricultural biotechnology and plays an important role in the in vitro regeneration of vegetable crops. Capsicum belongs to the Solanaceae Capsicum genus and is one of the main vegetables in my country. Capsicum L. has five cultivars: annual capsicum (Capsicum annuum), Chinese capsicum (Capsicum chinense), shrub capsicum (Capsicum frutescens), Capsicum baccatum and Capsicum pubescens. The seeds are black or nearly black, and the pulp is thick; the flowers are purple or light purple, with a white ring at the base, usually with a raised spot of a yellow nectary on the flower, and the plant is hairy. Capsicum pubescens and other capsicum cultivars cannot be pollinated to produce seeds, and there is reproductive isolat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/001
Inventor 汪志伟舒黄英朱丽蓉郝园园杨壮成善汉朱国鹏
Owner HAINAN UNIV
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