Application of ZmbHLH148 protein and coding gene thereof in regulation and control of plant drought resistance
A technology encoding gene and drought resistance, applied in the application field of ZmbHLH148 protein and its encoding gene in regulating plant drought resistance, can solve problems such as unclear mechanism of action, less corn, etc., achieve strong purpose, short time, improve The effect of plant drought resistance
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Embodiment 1
[0040] Example 1 Construction and detection of ZmbHLH148 gene carrier
[0041] using H 2 O and 20% PEG6000 screened the overexpression material of different genes when the phenotype of the germination stage, and accidentally found that the different lines of the ZmbHLH148 overexpression vector progeny all showed that the seed root was longer than the seed root length of the control LH244 under the PEG6000 treatment condition (such as image 3 , where A is shown in H 2 The germination of seeds under the treatment conditions of O and 20% PEG6000, picture B is in H 2 The root length statistical data under the O treatment condition, C picture is the root length statistical data under the 20%PEG6000 treatment condition), after querying, it is obtained that the carrier utilizes transgenic technology to overexpress the ZmbHLH148 gene from the corn genome (nucleotide sequence such as shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2). Its vector is construct...
Embodiment 2
[0045] Construction and identification of embodiment 2 transgenic plants
[0046] Transform the constructed pBCXUN vector containing the target gene into the Agrobacterium EHA105 strain, transfer it into the recipient material LH244 immature embryos under sterile conditions, and form callus after dedifferentiation, and then differentiate into tissues and organs. Transgenic corn seedlings. This seedling was marked as the T0 generation.
[0047] The transgenic material T0 was transplanted in the greenhouse of the West Campus of China Agricultural University, and the copy number of the marker bar gene in the transgenic lines was identified at the V3 stage of maize, and the T1 seeds of the transgenic lines with a lower copy number were harvested, and the T1 seeds were planted in At the Zhuozhou Experimental Station of China Agricultural University, when the seedlings reached the V3 stage, a homozygous individual plant of the bar gene was identified, and the homozygous self-crosse...
Embodiment 3
[0049] Example 3 Detection of germination phenotype of ZmbHLH148 overexpression material
[0050] Treat and sow under normal conditions, and germinate under 25°C culture conditions in an artificial climate chamber. The results show that LH244 has broken through the vermiculite to the surface on the 3rd day, while the transgenic line has not broken through the vermiculite (see figure 2 left), removed from the vermiculite on days 3 and 4, and photographed (see figure 2 In the middle panel, from top to bottom are the photos of the strains on the 3rd day and the 4th day respectively), it can be seen that the embryo length of the transgenic ZmbHLH148 strain is significantly shorter than that of the control material LH244. Statistics on the germ length of the transgenic material showed that the germ length of the transgenic material was significantly shorter than that of the control ( figure 2 right). It indicated that the gene was involved in the regulation of maize seed germi...
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