Paddy rice fragile straw controlling gene BC1 and use thereof
A gene and brittle stalk technology, applied in the field of rice BC1 gene, to achieve the effect of increasing content and quality, reducing environmental pollution, and improving material
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Embodiment 1
[0035] 1. Rice material
[0036] Rice (Oryza sativa ssp.) mutant Brittle Culm1-101, the original wild-type material is
[0037] "Shuangkezao" indica rice variety.
[0038] 2. Analyze and target groups
[0039] The homozygous bc1-101 mutant was crossed with the original wild-type variety C-Bao, and F 1 A total of 30,000 F 2 Individuals, and 7068 bc1-101 mutant individuals were selected as the mapping population. Take about 2 grams of young leaves from each plant at the seedling stage to extract DNA.
[0040] 3. Localization of BC1 gene by SSLP, RFLP, and CAPS markers
[0041] Using the Modified CTAB Method [9] Genomic DNA for gene mapping was extracted from rice leaves. About 100mg of rice leaves were taken, frozen in liquid nitrogen, ground into powder in a small mortar with a diameter of 5cm, transferred to a 1.5ml centrifuge tube to extract DNA, and the obtained DNA precipitate was dissolved in 100μl ultrapure water. 1 μl DNA sample was used for each SSLP and CAPS re...
Embodiment 2
[0048] Embodiment 2 transformed plants (rice is an example)
[0049] nh12 and aa06 are two plasmid clones from a random library used to sequence the BAC clone OSJNBa0036N23, which were joined by a common NheI site into a 5.8 kb (na18) fragment that includes the complete ORF region of BC1. At the same time, na18 was digested with BamHI to generate a 3.3kb fragment, which only contained the ORF of the first half of BC1. The 5.8kb and 3.3kb fragments were respectively cloned into the binary vector pCAMBIA1300, and the plasmids pCna18 and pCna181T for transformation were obtained (Fig. 5). These two plasmids were transformed into Agrobacterium tumefaciens strain LBA4404 by electric shock method to transform rice. Mature seeds of the bc1-101 mutant were dehulled and sterilized, and inoculated into a medium for inducing callus. After 3 weeks of culture, the callus was grown from the scutellum, and the vigorously growing, light yellow, relatively loose embryogenic callus was select...
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