Method for producing male sterile plants

一种雄性不育、植物的技术，应用在植物学设备和方法、生物化学设备和方法、植物产品等方向，能够解决破坏程序性细胞死亡、延迟等问题

Inactive Publication Date: 2011-11-23
COUNCIL OF SCI & IND RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This leads to a disruption of the normal tapetum cell death program and a delay in the induction of tapetum programmed cell death (PCD)

Method used

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  • Method for producing male sterile plants
  • Method for producing male sterile plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Isolation of encoding from Arabidopsis BECLIN 1 / ATG cDNA of 6 genes

[0091] Plant material: The Columbia ecotype (Col-O) of Arabidopsis thaliana was used in the experiments described here. Plants were grown on a complex soil mixture of vermiculite / sphagnum moss / perlite (1:1:1) in a growth chamber with a 16 h light / 8 h dark light cycle and a 23 °C day / 18 °C night temperature cycle .

[0092] Artificial induction of autophagy: The third and fourth rosette leaves of 3-week-old plants were isolated and floated on deionized water in a Petri dish, adaxial side up. The leaves were incubated for 48 hours at 22±1°C in the dark.

[0093] plant BECLIN 1 / ATG6 Cloning of genes: Total RNA was extracted from autophagy-induced Arabidopsis leaves using TRI reagent (Sigma). Using NanoDrop ? ND-1000 UV-Vis spectrophotometer measures the amount of total RNA. RNA quality was checked by visualizing rRNA in ethidium bromide-stained agarose gels under UV light. 10 µg of total R...

Embodiment 2

[0095] Tapetum-specific promoters and plants BECLIN 1 / ATG Construction of chimeric gene fusions between 6

[0096] In early stages, the tapetum-specific 1 kb BamH1 / Xba1 promoter associates with the 1.5 kb Xba1 / Sac1 plant BECLIN1Gene and 250 bp Sac1 / EcoR1 Nos terminator fused in BamH1 / EcoR1 Sk+ cloning vector.

[0097] The entire expression cassette containing the fragment BamH1 / EcoR1 was further subcloned into the binary vector pBI101. The resulting pBI101 carrying the expression cassette was introduced into Agrobacterium tumefaciens strain LBA4404 according to a modified protocol (Cangelosi et al., 1991).

Embodiment 3

[0099] Transformation of Tobacco Plants

[0100] As described in Example 2, Petit Havana tobacco ( Nicotina tabacum cv. Petit Havana ), using the protocol described by Horsch et al. in 1985.

[0101] Briefly, a single isolated colony of Agrobacterium tumefaciens LBA 4404 containing the binary vector with the expression cassette described above was inoculated into a drug containing the antibiotics streptomycin (250 mg / ml), rifampicin (50 mg / ml) and kana Mycin (100mg / ml) in YEP medium and allowed to grow (200 rpm, overnight, 28°C). 50 μl of overnight culture was diluted to 100 ml in YEP medium and allowed to grow to OD 600 reached 0.8. Cells were recovered by centrifugation in an SS34 rotator (5,000 rpm, 10 min, 4°C). Suspend the pellet to OD in co-cultivation medium (MS salts, 2% glucose, 10 mM MES and 100 mM acetosyrengone, pH 5.6) 600 0.6. Tobacco leaf discs were co-cultured with Agrobacterium tumefaciens for 2 days in the dark. After co-cultivation, the leaf disc...

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Abstract

The present invention relates to a method for producing male sterile plants. It involves selective killing of reproductive cells in plants by using an autophagy related gene plant BECLIN I / ATG6. An expression cassette comprising plant BECLIN 1 / ATG6, regulated by a tapetum specific promoter can induce killing of tapetum cells. A particular area of interest is transforming a plant with said genetic construct and expression of the plant BECLIN I gene in tapetum at early stage of anther development to cause early collapse of tapetum to produce nonviable pollen, thus imparting male sterility in plants.

Description

technical field [0001] The present invention relates to a method of producing male sterile plants. Specifically, the invention relates to the use of plants comprising Beclin 1 Transforming plants with the gene construct for the gene and expressing the gene in the anther tapetum produces male sterile transgenic plants. Male sterile plants are useful in the production of hybrid plants by sexual crossing. The development of hybrid varieties is highly desirable because of their increased heterosis (hybrid vigor or heterosis) resulting in generally increased yields. Background technique [0002] Hybrid plants have become increasingly important in a variety of commercial food crops worldwide. Hybrid plants have the advantage of higher yield, better quality and stress resistance than their parents due to heterosis. Crop uniformity is another advantage of hybrid plants when the parents are homozygous; this leads to improved crop management. Hybrid seeds are therefore commercial...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
CPCC12N15/8289
Inventor S.V.萨万特R.图利S.P.辛
Owner COUNCIL OF SCI & IND RES
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