Method for cultivating actinidia arguta anther into haplobiont

A technology of kiwi fruit and haploid, which is applied in the fields of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of long cycle and low efficiency of hybrid breeding, achieve simple operation, improve selection and breeding materials Effect

Active Publication Date: 2017-11-03
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] Actinidia jujube is generally tetraploid, with 116 chromosomes (2n=116, 4X), highly heterozygous genes, and rich genetic diversity. The traditional hybrid bree...

Method used

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  • Method for cultivating actinidia arguta anther into haplobiont
  • Method for cultivating actinidia arguta anther into haplobiont
  • Method for cultivating actinidia arguta anther into haplobiont

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Experimental program
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Embodiment 1

[0033] 1) Experimental material: Select 'Kuilu' plants that grow robustly and are free from diseases and insect pests. From 9:00 to 11:00 in the morning on a sunny day, the immature flower buds of "Kuilu" were collected, and the microspore development period was observed by the acetic acid magenta staining method. use;

[0034] 2) Low temperature treatment: put the collected flower buds in an environment of 2°C for pretreatment for 1 day. After the pretreatment, flower buds in the mononuclear marginal stage are selected respectively, and after disinfection and sterilization, the anthers are stripped. Anthers were inoculated on callus induction medium. 5-7 anthers were inoculated in each bottle, 10 bottles were inoculated for each treatment, and repeated 3 times. Cultivate in the dark for 20 days at 23°C;

[0035] 3) Induction of plant regeneration: when the induced callus grows to a diameter of 3-5 mm, it is transferred to the plant differentiation medium of Actinidia jujub...

Embodiment 2

[0039] 1) Experimental material: Select 'Kuilu' plants that grow robustly and are free from diseases and insect pests. From 9:00 to 11:00 in the morning on a sunny day, the immature flower buds of "Kuilu" were collected, and the microspore development period was observed by the acetic acid magenta staining method. use;

[0040] 2) Low temperature treatment: put the collected flower buds in an environment of 4°C for pretreatment, Process 1d, 3d, 5d, 7d, 10d After the pretreatment, the flower buds in the mononuclear stage were selected, rinsed with tap water for 30 minutes, and under aseptic conditions, the flower buds were sterilized with 70wt% alcohol for 30 seconds, rinsed with sterile distilled water for 3 to 4 times, and washed with 0.09wt% HgCl 2 The solution was sterilized for 5 minutes, rinsed with sterile distilled water for 3 to 4 times, and the anthers were stripped off. The anthers were inoculated on callus induction medium (MS+2,4-D 0.5 mg / L+6-BA 1.0 mg / L+sucrose ...

Embodiment 3

[0049] 1) Experimental material: Select 'Kuilu' plants that grow robustly and are free from diseases and insect pests. From 9:00 to 11:00 in the morning on a sunny day, the immature flower buds of "Kuilu" were collected, and the development of microspores was observed by acetic acid magenta staining. Tetrad, single-core early, single-core to the side inflorescence apical bud , collecting the flower buds at the top of the inflorescence where the megaspore development is in the uninucleate marginal stage for several days, and set aside;

[0050] 2) Low-temperature treatment: put the collected flower buds in an environment of 2°C for pretreatment for 5 days. After the pretreatment, select the flower buds in the single-nucleus marginalization stage, wash them with tap water for 30 minutes, and wash the flower buds with 75wt% alcohol under aseptic conditions. Disinfect for 30s, rinse with sterile distilled water 3 to 4 times, and use 0.12wt% HgCl 2 The solution was sterilized f...

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Abstract

The invention provides a method for cultivating actinidia arguta anther into haplobiont. The method comprises the following steps: (A) collecting flower buds developed by actinidia arguta microspores, carrying out low-temperature refrigeration treatment for 1-10 days, and peeling anther; and (B) putting the anther in a callus tissue induction medium for carrying out dark culture for 20 days or more, culturing the anther for 40 days or more in a plant differentiation medium, and finally transferring the anther into a rooting medium for culturing and rooting to obtain the haplobiont. Through the method for cultivating actinidia arguta anther into haplobiont of the embodiment, multiple homozygous breeding materials are provided for breeding of actinidia arguta; selection and breeding materials are provided; meanwhile, an ideal receptor material is provided for the basic theoretical research; compared with the manners such as an in-vitro microspore culturing manner and a somatic chromosome eliminating manner, the anther culturing method is simple and convenient to operate; a method of artificially culturing in-vitro anther is an effective method for obtaining haplobiont of actinidia arguta; the method is extremely worthy of extensive popularization and application.

Description

technical field [0001] The invention relates to the field of kiwifruit cultivation, in particular to a method for cultivating anthers of kiwifruit into haploid plants. Background technique [0002] Jujube kiwifruit, also known as soft jujube, kiwi pear, vine melon, vine pear, belongs to Actinidiaceae, and kiwifruit is a large deciduous vine plant. The fruit is small, smooth, and the whole fruit is edible. It is one of the most cold-resistant species in the genus Actinidia. one. Jujube kiwifruit enjoys the reputation of "the king of fruits". It is favored by consumers for its excellent quality, flavor, rich nutritional content, high medical and health care effects, and good processing suitability. Actinidia jujube is a promising small berry tree species in many regions of the world, especially in cold regions. The surface of kiwi fruit is smooth and hairless, with a lot of juice and good flavor. The pulp is rich in vitamins and amino acids, and the content of vitamin C is p...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 秦红艳王广富艾军孙丹范书田杨义明许培磊刘迎雪赵滢王振兴张宝香李昌禹
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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