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Application of PLD alpha1 gene in aspects of increasing crop drought resistance and seed production

A technology of drought resistance and genetics, applied in the field of plant molecular breeding and biology, can solve the problems of limitations and unclear effects of PLD, and achieve the effects of enhancing biological yield, reducing water transpiration, improving water retention capacity and drought resistance

Inactive Publication Date: 2014-05-07
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the improvement of plant stress resistance of PLDα1 is mostly limited to the model organism Arabidopsis, and the role of PLD in crop drought resistance is still unclear

Method used

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  • Application of PLD alpha1 gene in aspects of increasing crop drought resistance and seed production
  • Application of PLD alpha1 gene in aspects of increasing crop drought resistance and seed production
  • Application of PLD alpha1 gene in aspects of increasing crop drought resistance and seed production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Rapeseed genetic transformation (rape transgenic means-dark light cultivation method)

[0032] Seed sterilization and germination: After soaking the seeds with 75% alcohol for 1 min, rinse the seeds with sterile water. Then use 0.15% HgCl 2 (containing 0.1% (v / v) Tween20) for 15 minutes, then rinsed with sterile water for 3-5 times; then sow the seeds in M 0 In the culture medium, 18 seeds were evenly sown per dish (3 cm in diameter). Put the petri dish into a sterile plastic culture box, and culture it at 25°C for 5 days under dark and light conditions.

[0033] Cultivation of Agrobacterium: Pick a single colony of Agrobacterium (GV3101, a strain containing the AtPLDα1 gene plasmid, purchased through a commercial channel) and put it in LB liquid medium (containing 50 mg / L kanamycin) for 1-2 days. The best infection OD value is 0.6-0.8. Use a spectrophotometer to detect the concentration of the bacterial solution under the condition of 600nM light abso...

Embodiment 2

[0043] Example 2: Identification and expression of PLDα1 transgenic plants

[0044] (1) Identification of PLDα1 transgenic plants

[0045] Plants transformed with Agrobacterium in M 4 After the roots grow in the culture medium, take an appropriate amount of leaves (preferably newly grown leaves far away from the culture medium to prevent Agrobacterium contamination) to extract DNA, and use the primer KatI-p / PLDα1-A to perform DNA extraction on the leaves. PCR detection, while the plasmid was used as a positive control, and the DNA of WT plants was used as a negative control, to identify whether the AtPLDα1 gene was successfully transferred into the genome of the regenerated plants resistant to kanamycin. It can be seen from the results that most of the plants can amplify a fragment with the same size as the PCR product of the plasmid as a template, while a few lines and the control wild-type WT do not amplify the current fragment, which shows that the culture in dark light T...

Embodiment 3

[0061] Example 3: AtPLDα1-OE plants enhance drought tolerance and seed yield

[0062] (1) The stomata of AtPLDα1-OE plants are more sensitive to ABA

[0063] In order to test the effect of AtPLDα1-OE plants on water use, the transgenic plants of Westar and Zhongshuang 9 were planted in soil and grown in a growth chamber. Use a hole punch to remove about 0.5 cm from the fully expanded leaves of the 23-day leaf year 2 The leaf discs were placed in the buffer that induces stomatal opening (under light conditions) and incubated for 1 h, and the opening of the stomata was observed under a microscope. When about 90% of the stomata are open, add 50 μM ABA to the buffer and continue to incubate for 30 min, and observe the change of stomatal conductance under a microscope. The results showed that the stomatal diameter of AtPLDα1-OE plants was not significantly different from that of WT plants when the stomata were induced to open by light. However, after adding ABA for 30 min, the s...

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Abstract

The invention belongs to the fields of plant molecular breeding and biotechnology and discloses application of a PLD alpha1 gene in aspects of increasing crop drought resistance and seed production. The nucleotide sequence of the PLD alpha1 gene is shown as SEQ ID NO:1, or the similarity of the nucleotide sequence of the PLD alpha1 gene is more than or equal to 70% compared with the nucleotide sequence shown as SEQ ID NO:1. The invention also discloses a method for increasing rape drought resistance and rape seed production. Experiments show that rape with stoma-specific overexpressed AtPLDalpha1 can effectively regulate stomas of plants to be closed under drought stress, so that moisture transpiration of leaves is reduced, and the water retaining capacity and the drought resistance of the plants are improved; the genetically modified rape plant has the characteristics that the flowering phase is postponed, vegetative growth is vigorous, the silique quantity is high, and the main inflorescence and the plant height of the rape are remarkably higher than those of the wild type rape when being suffered from an arid season in an early phase or an intermediate phase under a natural growth condition in the fields.

Description

technical field [0001] The invention belongs to the field of plant molecular breeding and biotechnology, and specifically relates to the application of PLDα1 gene in increasing crop drought resistance and seed yield and a method for increasing crop drought resistance and seed yield. Background technique [0002] With the change of climate and the increase of arid area year by year, drought has become one of the main factors restricting crop yield, and irrigation agriculture has become an important measure to improve agricultural production. According to statistics, modern agricultural irrigation water accounts for about 70% of fresh water resources, and the efficient use of water resources for agricultural production has become the focus of modern botanists. People hope to increase agricultural output by increasing the water used for plant irrigation. However, the fresh water resources on the earth are gradually decreasing. Improving the drought resistance and water use effi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55C12N15/84A01H5/00A01H4/00
Inventor 鲁少平王学敏洪月云
Owner HUAZHONG AGRI UNIV
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