A Molecular Marker Co-segregated with Rice Photothermosensitive Genic Male Sterility Trait and Its Application
A nuclear-sensitive and male-sterile technology, applied in the field of plant molecular biology, can solve problems such as little understanding of molecular basis
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Embodiment 1
[0042] Phenotypic and Genetic Analysis of Rice light tms4430 embodiment 1 Example mutation PTSGMS
[0043] Screening 1) tms4430 mutant
[0044] June 2013 93-11 10 kilograms radiation seeds get M0-generation cobalt-60. The radiation seeds planted in experimental plots Lingao County, Hainan Province, mature sub-species of plant closing, M1 generation materials total of about 6,500. The spring of 2014, many election of 3617 the amount of seed planted into the material M1 generation lines, each line 50 plant species. Respectively tillering, booting, heading, flowering, grain filling screening plant type, spike type, fertility, yield and other types of mutants were preserved and closed. Number 4430 in which the system performance is found that a mutant male sterility, is named tms4430.
[0045] Phenotypic 2) tms4430 Mutants
[0046] In cultivation conditions lingao positive quarter with iodine - potassium iodide solution (0.6% KI, 0.3% I 2 , W / w) of different grass pollen were staine...
Embodiment 2
[0049] Preliminary positioning light TMS4430 temperature sensitive male sterility gene of Example 2
[0050] A mixing tank using extreme trait analysis (bulked segregant analysis) and recessive population analysis (recessive-class analysis) of TMS4430 gene targeting. In the present MH63 parent tms4430 we constructed a mutant of F2 population, isolated in 169 sterile plants. Using the 301 pairs of SSR uniformly distributed on the screening of chromosomal markers 12 markers polymorphic between 93-11 and MH63, screened 64 pairs of polymorphic SSR markers, low polymorphism showed clear 63 and 93-11 recovery smaller difference in the genomic sequence. The F2 population were constructed fertile plants and sterile plants mixed pool of genes, polymorphic markers with the 64 pairs of the gene pool of the fertile and sterile gene pool analysis. The results indicated the presence of linkage SSR markers RM243 and RM023 and the mutant phenotype tms4430 on chromosome 1. Genotyping by-sterile pl...
Embodiment 3
[0051] Example 3 Rice Fine Mapping light of temperature sensitive male sterility gene TMS4430
[0052] Based on the above results of the rough positioning, recessive population analysis (recessive-class analysis) for fine positioning. Using markers linked heteroaryl selected tms4430 F2 population developed a laminated F3 plant population, comprising 3827 plant, wherein the isolated 952 sterile plants. Meanwhile, between the marks and the RM10544 RM10495 further developed three polymorphic markers. Wherein the primer for detecting the molecular marker Indel14270 as shown in SEQ ID NO.1-2; molecular marker for detecting Indel14340 primer as shown in SEQ ID NO.3-4; means for detecting the primer shown in SEQ M3193-6 FIG IDNO.5-7.
[0053] Using these markers individually detected F3 populations isolated genotype sterile plants, found Indel14270 sterile plants only mark an expanded band of 305bp ( image 3 ); Indel14340 mark only in the expanded band of 132bp ( Figure 4 ); Only the numer...
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