Cucumber CsGL2-LIKE gene and application thereof in regulating partial abortion of male flowers

A cucumber, genetic technology, applied in the direction of application, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2019-11-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report about the involvement of HD-ZIP IV in cucumber male sterility

Method used

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  • Cucumber CsGL2-LIKE gene and application thereof in regulating partial abortion of male flowers
  • Cucumber CsGL2-LIKE gene and application thereof in regulating partial abortion of male flowers
  • Cucumber CsGL2-LIKE gene and application thereof in regulating partial abortion of male flowers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Cloning of gene CsGL2-LIKE related to partial abortion of cucumber male flowers

[0035] 1. Materials

[0036] Cucumber 3461 (the variety selected in our laboratory) (Liu et al., 2015; Silencing of the gibberellin receptor homolog, CsGID1a, affects locule formation in cucumber (Cucumis sativus) fruit. NEW PHYTOLOGIST, 210, 551-563.) was used as the test material , the breed belongs to the hermaphroditic line. The cucumber seeds are sown in the solar greenhouse, and the field management is the same as the normal cultivation management. When the male flowers appeared on the plants, the male flower buds were quick-frozen in liquid nitrogen and stored in a -80°C refrigerator for future use.

[0037] 2. Total RNA extraction and cDNA synthesis

[0038] (1) Extraction of total RNA

[0039] The total RNA of cucumber male flower buds was extracted with a plant total RNA extraction kit (purchased from Beijing Huayueyang Biotechnology Co., Ltd.), and the residual DNA...

Embodiment 2

[0067] Embodiment 2, the acquisition of transgenic plants and its detection

[0068] 1. Genetic transformation of cucumber

[0069] (1) Transformation of Agrobacterium with the recombinant expression vector prepared in Example 1: Turn on the Biorad electroporator to preheat, and at the same time place the electroporation cup on ice to precool, and put it into an ultra-clean workbench for ultraviolet sterilization for 30 minutes to sterilize. Add the constructed CsGL2-LIKE-pCAMBIA1008 plasmid (interference expression vector constructed for Example 1 step 5) and pCAMBIA1008 empty plasmid 500ng into 100 μl of C58 Agrobacterium competent cells respectively, flick and mix well (Agrobacterium competent Place the cells on ice in advance to thaw) Add the above-mentioned mixed competent state to the bottom of the electroporation cup, then close the lid and place it on the seat of the electroporation apparatus for electric shock. After the end, add 300ul of liquid YEB medium without an...

Embodiment 3

[0094] Embodiment 3, phenotypic identification of transgenic plants

[0095] Eight transgenic lines were obtained by constructing CsGL2-LIKE RNAi interference vectors to transform cucumbers, among which the lines with more serious interference of CsGL2-LIKE RNAi expression were #5, #7, #21 and #25 (see Figure 4 ). Through careful observation of the transgenic plants, it was found that the fruit thorn density and size of the transgenic plants had no change compared with the wild type (see Figure 5 ). However, in the development of male flowers, there are significant differences between the transgenic lines and the wild-type lines, as follows: (1) The male flower opening time of the interference lines is significantly delayed compared with that of the wild type (see Figure 6A ), the internal structure is not fully developed compared with the wild type (see Figure 6B ); the fresh weight of the male flower was about 50% of the wild type on the day of opening (see Figure 6...

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Abstract

The invention provides a cucumber CsGL2-LIKE gene and application thereof in regulating the partial abortion of male flowers. The nucleotide sequence of the cucumber CsGL2-LIKE gene is shown as SEQ IDNO. 2, and the amino acid sequence of an encoded protein of the gene is shown as SEQ ID NO. 1. An interference vector of the CsGL2-LIKE gene is constructed and then transformed into cucumber by an agrobacterium-mediated transformation technology; it is proved that the down-regulation of the CsGL2-LIKE gene causes the change of the shape and pollen viability of cucumber stamens, resulting in the partial abortion of the male flowers of transgenic plants, the decline of pollen viability and the weakening of the elongation ability of pollen tubes; however, female flowers are not affected, it is shown that the function of the CsGL2-LIKE gene is related to the partial abortion of the cucumber male flowers, and it is indicated that the gene can have an important improvement potential in cucumberbreeding and quality improvement.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and specifically relates to cucumber CsGL2-LIKE gene, its encoded protein and application. Background technique [0002] Plant male sterility refers to the abnormal development of male gametes and normal development of female gametes, which can accept foreign pollen for normal pollination. According to the difference of inheritance mode, plant male sterility is mainly divided into nuclear sterility (Genic male sterility, GMS) and cytoplasmic male sterility (cytoplasmic male sterility, CMS). According to the stage of abortion and the characteristics of anther histocytology, male sterility can be further divided into sporophytic male-sterility with abnormal development of tapetum and pollen, and gametophytic male-sterility with abnormal development of microspores or pollen grains. There are two forms of gametophytic male-sterility. Male sterility has always been a hot spot in breeding. Fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/02A01H6/34C12N15/11
CPCC07K14/415C12N15/827C12N15/8289
Inventor 刘兴旺任华中蔡艳玲董明明张亚琦
Owner CHINA AGRI UNIV
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