Rice albino-transforming green leaf gene val1 and its encoded protein and application
A rice albino technology, applied in the field of genetics, can solve problems such as inconvenient removal of impurities, difficulties, and poor traits
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Embodiment 1
[0028] Example 1. Obtaining and Morphological Observation of Rice Albino-transformed Green Leaf Mutant val1
[0029] A genetically stable albino-to-green mutant of rice leaf color was obtained by using ethyl methanesulfonate (EMS) to mutate the self-fertile excellent restorer line Jinhui 10, named val1.
[0030] In wild type (WT), plant leaves are green throughout the growth period except at the late stage of maturation. In the mutant val1, albino leaves appeared early in the seedlings ( figure 1 in A). From the late seedling stage to tillering stage, the leaves gradually turn green and the edges of the leaves are albino, showing a mixed phenotype ( figure 1 in B). At the heading stage, the leaves were almost gray-green, and only a few leaves showed marginal albinism ( figure 1 in C). Therefore, the older the leaf, the smaller the area of albinism. These results suggest that the val1 phenotype is regulated by reproductive progression. In addition, at the seedling a...
Embodiment 2
[0031] Molecular identification of embodiment 2, VAL1 gene
[0032] F 1 Generation plants, all the leaves are normal green, and then 3853 F 2 In the generation population, two phenotypes of mutant leaves and normal leaves were separated according to the traits of albino to green leaves, and 2885 normal plants and 968 mutant plants were isolated. The segregation ratio of 3:1 indicated that the mutant trait was controlled by a pair of recessive single genes.
[0033] Fine mapping: The VAL1 gene was preliminarily mapped in the range of 171kb between M22 and ID27 on chromosome 8. According to the published sequence of indica rice variety 93-11, markers were further screened and developed in this interval, and finally the VAL1 gene was fine-mapped between the simple repeat marker SSR8-1 and the insertion / deletion marker ID30 (sequence shown in Table 1). The distance is 29.69kb. There are 4 annotated genes in this interval (http: / / www.gramene.org / ).
[0034] Table 1, 2 pairs of...
Embodiment 3
[0037] Example 3, the expression pattern of VAL1 and the subcellular localization of VAL1 protein
[0038] In order to determine the expression pattern of VAL1, the primers in Table 2 were used for real-time fluorescence quantitative analysis. With Actin as the internal reference, the reaction system is: add 2 μL of cDNA template, 2 μL of primers, 12.5 μL of SYBR Green fluorescent dye and 8.5 μL of RNase-free H in a 25 μL reaction system 2 O, fluorescent quantitative amplification was performed on a Bio-rad fluorescent quantitative PCR instrument; the amplification conditions were: pre-denaturation at 94°C for 2 minutes; denaturation at 94°C for 30 seconds, renaturation at 56°C for 30 seconds, extension at 72°C for 1 minute, 40 Cycling; the final extension at 72°C for 10 minutes, and then use CFX-Manager software for data collection and processing, the results are as follows image 3 shown. Depend on image 3 It can be seen that VAL1 is expressed in various tissues, includi...
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