Tissue culture method taking masson pine in-vitro mature embryo as explant

A tissue culture and masson pine technology, which is applied in the fields of botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of only slight elongation or stop growth, unable to meet the requirements of production, difficult tissue culture of masson pine, etc. , to achieve consistent and robust growth, fast start-up, and high rooting rate

Inactive Publication Date: 2013-10-09
NANJING FORESTRY UNIV
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the tissue culture of Pine massoniana is difficult, and the induced buds are not easy to differentiate and elongate, and the rooting rate of seedlings is extremely low.
Huang Jianqiu, Wei Zhiming, etc. conducted somatic embryo induction on mature embryos of Pinus massoniana and obtained regenerated plants. A total of 27 cotyledon embryos were induced, and finally only 2 complete plantlets were produced, and the rest could only slightly elongate or stop growing. The frequency of embryo transformation into small plants is only 7.4%; Zhang Yu’s tissue culture of Masson pine has a certain rate of bud induction, but the rooting rate is only 70%, which cannot meet the production requirements

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The tissue culture method using the isolated mature embryo of Pinus massoniana of the present embodiment as an explant may further comprise the steps:

[0021] The first step is to collect materials: take the mature seeds of Pinus massoniana, soak them overnight, rinse them under running water, and peel off the outer seed shell with tweezers.

[0022] The second step, material disinfection: the shelled seeds are sterilized with 75% ethanol for 30s in the ultra-clean workbench, then sterilized with 0.1% mercury liter for 10-15min, and finally rinsed with sterile water for 3-5 times.

[0023] The third step, inoculation and adventitious bud induction: In the ultra-clean workbench, use the method of conventional aseptic operation, use a scalpel and tweezers to peel off the seed embryos, inoculate them horizontally on the induction medium, and be careful when peeling off the seed embryos Be careful to avoid damage to the embryos as much as possible. The composition of the i...

Embodiment 2

[0032] The difference from the above examples is that the isolated mature embryos of Pinus massoniana are inoculated on the induction medium and cultured for about 20 days. Divided into DCR conventional basic medium, 6-benzylpurine 2.0 mg / L, naphthalene acetic acid 0.15 mg / L, sucrose 30 g / L; transfer the culture to the differentiation medium, cultivate for several days, and wait for the small buds to differentiate After that, the small buds can be inoculated on the bud elongation medium to elongate the buds, cultivated for 30-40 days, and when the young shoots grow to the height required by the conventional rooting procedure, enter the next step, wherein the components of the differentiation medium It is: DCR conventional basic medium, 6-benzylpurine 0.5 mg / L, indolebutyric acid 0.05 mg / L, sucrose 30 g / L, agar 6.0 g / L, and the composition of shoot elongation medium is: DCR Conventional basal medium, naphthalene acetic acid 0.1 mg / liter, sucrose 30 g / liter, this medium promotes...

Embodiment 3

[0034]The difference from the above examples is that the components of the induction medium are DCR conventional minimal medium, 1.0 mg / liter of 6-benzylpurine, 0.10 mg / liter of naphthaleneacetic acid, and 30 g / liter of sucrose; The components are: DCR conventional basic medium, 0.5 mg / L 6-benzylpurine, 0.05 mg / L indolebutyric acid, 30 g / L sucrose, 5.7 g / L agar; the components of the shoot elongation medium are : DCR conventional basic medium, indole butyric acid 0.05 mg / liter, sucrose 30 grams / liter; the components of the rooting medium are: 1 / 2DCR conventional basic medium, indolebutyric acid 2.0 mg / liter, sucrose 20 grams / Lift.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a tissue culture method taking a masson pine in-vitro mature embryo as an explant. The tissue culture method comprises the following steps of: adventitious bud induction, namely, inoculating the masson pine in-vitro mature embryo on an inducing medium, culturing for about 20 days, and after inducing out bud primordia from a culture, carrying out the next step; differentiation and elongation of the buds: namely, (1) transferring the obtained culture to a differential medium, culturing for several days, after forming budlets through differentiation, inoculating the budlets on a bud elongation culture medium for carrying out bud elongation, culturing for 30-40 days, and when plantlets grow to heights required for a conventional rooting procedure, carrying out the next step; and rooting: namely, taking down the obtained plantlets obtained in the step (2) from the roots, transferring the plantlets to a conventional rooting medium, after culturing for 28-35 days, white short roots begin to occur at the roots of the plantlets, at the moment, transferring the rooting plantlets to a root elongation culture medium, culturing for about 30 days, then transplanting and finally growing into normal masson pine plants.

Description

technical field [0001] The invention relates to a tissue culture method using isolated mature embryos of pinus massoniana as explants, and belongs to the technical field of woody plant tissue culture. Background technique [0002] Masson pine is native to my country. It is one of the most important tree species in the provinces south of the Qinling Mountains for greening barren hills, papermaking raw material forests, resin mining forests, and natural landscape forests. It is the most widely distributed tree species in my country. It grows rapidly, is highly productive and adaptable. Good mechanical properties and other characteristics, it is also widely used in construction, pit wood, sleepers, bridges and other projects, and can also be used as raw materials for fiberboard, particleboard, and plywood industries, and has important economic value. At present, the improved varieties of masson pine are mainly sexually propagated in seed orchards and mother forests. However, ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 季孔庶王金玲王潘潘阮倩倩
Owner NANJING FORESTRY UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap