A rice white spot leaf control gene wlml1 and its encoded protein and application
A white-spotted leaf and protein technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as affecting rice photosynthetic efficiency, crop yield reduction, and rice plant death.
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Embodiment 1
[0037] 1. Rice material:
[0038] The original wild type of rice (Oryza sativa L.) leaf color mutant wlml1 (white and lesion mimic leaf1) is the indica variety TN1 (Taichung Native 1).
[0039] The process of obtaining the rice (Oryza sativa L.) leaf color mutant wlml1 is as follows:
[0040] The seeds of the indica rice variety TN1 were mutated by EMS and planted in the field. The seeds were collected and planted continuously. From M 2 A white and lesion mimicleaf mutant was isolated from the generation, and it was tentatively named wlml1 (white and lesion mimicleaf1). The mutant is self-crossed for multiple generations to obtain a mutant plant that can be stably inherited.
[0041] The mutant material wlml1 and the wild-type material TN1 were reciprocally crossed, that is, the mutant wlml1 was used as the female parent and TN1 was used as the male parent for crossing; at the same time, TN1 was used as the female parent and wlml1 was used as the male parent for crossing. E...
Embodiment 2
[0081] Plant Transformation:
[0082] Using wild-type genomic DNA as a template, the target gene was cloned by PCR. The clone covered the entire genomic region of the ORF of the target gene, including the 2.1 kb sequence upstream of ATG and the 1.1 kb sequence downstream of TAG. After electrophoresis detection and gel recovery, the target gene was connected to the pCAMBIA1300 plasmid digested by EcoRI and XbalI by homologous recombination. The plasmid was transformed into Agrobacterium tumefaciens strain EHA105 by electric shock method, and then the rice callus was transformed by Agrobacterium tumefaciens. We used the callus induced by the mutant immature embryos, and after 3 weeks of culture in the induction medium, we selected the vigorously growing callus as the recipient of transformation. The rice callus was infected with the EHA105 strain containing the binary plasmid vector, co-cultivated in the dark at 25°C for 3 days, and cultured on a selection medium containing 40 ...
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