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Rice anther development regulation gene EDT1 and application thereof

A technology for EDT1 and gene regulation, applied in the application field of rice anther development-related protein EDT1 and its coding gene, can solve the problem of less research on ACL, and achieve the effect of strong stress resistance and wide adaptability

Active Publication Date: 2021-02-09
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, some molecules derived from acetyl-CoA (such as polyester polyhydroxybutyrate) have high industrial value[11] in the pharmaceutical and chemical industries, and how to control the production of acetyl-CoA Carbon flux is an important topic for many biotechnology applications[12], however ACL is less studied in other aspects

Method used

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  • Rice anther development regulation gene EDT1 and application thereof
  • Rice anther development regulation gene EDT1 and application thereof
  • Rice anther development regulation gene EDT1 and application thereof

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Experimental program
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Effect test

Embodiment 1e

[0055] Phenotype observation of embodiment 1edt1 mutant and map-position cloning of EDT1 gene

[0056] (1) The phenotype of the edt1 mutant was observed to clarify its mutation type

[0057] The edt1 mutant was obtained from the indica rice variety IR64 through tissue culture. After field observation, we found that the defective phenotype of the mutant (short plant height, short and white anthers) existed stably ( figure 1 , A, plant phenotypes of wild type and edt1 mutant after heading; B, spikelets of wild type and edt1 mutant after removing palea and lemma; C, anthers of wild type and edt1 at heading stage; D, wild Anthers of type and edt1 use iodine-potassium iodide (I 2 -KI) solution staining), the mutants were ultimately completely sterile because they could not form normally fertile pollen grains. After acetic acid magenta staining, it was found that in the edt1 mutant, after meiosis, microspores were released from the tetrad as in the wild type, but then developmenta...

Embodiment 2

[0066] Example 2 EDT1 participates in the degradation process of tapetum in anther

[0067] We analyzed the anther walls of wild-type and edt1 mutant anthers by TUNEL assay and comet assay, respectively. The experimental results found that in the wild-type tapetum cells, TUNEL positive signal began to appear at stage 8 (during meiosis), and a strong TUNEL signal was detected during stage 9 (microspore stage). In the mutant, we detected TUNEL positive signal at the 7th stage, and the TUNEL signal was the strongest at the 8th stage, and then the signal gradually disappeared ( Figure 4 , Detection of anther DNA fragmentation from stage 5 (S5) to stage 10 (S10) of wild-type (A-C, G-I) and edt1 mutants (D-F, J-L). Red fluorescence indicates propidium iodide (PI)-stained nuclei. Yellow fluorescence is an overlay of TUNEL positive nuclear staining (green) and propidium iodide (red). Arrows in F, G, H, J show TUNEL positive signal in tapetum cells). The comet experiment quantifie...

Embodiment 3

[0068] Example 3 EDT1 affects oxidation-reduction balance and energy metabolism in anthers

[0069] Staining experiments on anthers by NBT dyes found that in edt1, O 2 .- Consistently high levels of ( Figure 6 A). Subsequently, we observed the paraffin sections of anthers at the ninth stage, and found that O 2 .- Mainly present in the tapetum cells ( Figure 6 B-C). For the quantitative detection of O 2 .- Through WST treatment, quantitative detection found that O in S10 and S11 stages in edt1 anthers 2 .- The deletion was significantly higher than that of the wild type ( Figure 6 D). According to the above experimental results, we speculate that the tapetum degradation mode of edt1 is different from that of the wild type, and its degradation may be caused by insufficient energy in cells. It was found by high performance liquid chromatography that the amount of ATP in the edt1 mutant was significantly lower than that of the wild type at the 8th and 9th periods; a...

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Abstract

The invention discloses a rice anther development regulation gene EDT1 and application thereof. The DNA sequence of the rice anther development regulation gene EDT1 is shown as SEQ ID NO.1. The rice anther development regulation gene EDT1 is obtained by researching a male sterility mutant edt1. The rice anther development regulation gene EDT1 can provide excellent provenance to cultivate high-quality rice varieties with high yield, high stress resistance and wide adaptability and has important application value in plant breeding.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering and relates to the application of a rice anther development-related protein EDT1 and its coding gene. [0002] technical background [0003] Rice fertility is a key factor affecting rice yield. In flowering plants, male gametogenesis is an important biological process, and it is also a growth process commonly experienced by higher plants. In the study of male development in plants, anthers are important research objects, and the external environment or abnormal changes in intracellular substances at any stage of anther development may lead to male sterility. During pollen development, the innermost layer of the anther wall, the tapetum, which can directly contact and interact with microspores, plays a crucial role. In the early days, people used the experimental method of cytology to study the specific function of the tapetum, and described it in relative detail. In recent years...

Claims

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Application Information

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IPC IPC(8): C12N15/60C12N9/88C12N15/82A01H5/02A01H6/46
CPCC12N9/88C12N15/827C12N15/8289
Inventor 万建民赵志刚柏文婷洪骏余晓文王超龙江玲田云录刘喜刘世家陈亮明
Owner NANJING AGRICULTURAL UNIVERSITY
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