Cloning and application of tomato velvet regulating gene
A technology for regulating genes and hairs, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as unclear regulation mechanisms.
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Embodiment 1
[0022] Example 1 Segregation Population Construction
[0023] The tomato materials used in this study include the conventional cultivar Ailsa Craig (AC), the Lanata (Ln) mutant LA3127, and the introgression line IL3 produced by the hybridization of Pennelli tomato (S. pennellii) / cultivated tomato (S. lycopersicum) M82. -2, LA1589 is gooseberry tomato, and the mutant was cited from the Tomato Genetic Resource Center (TGRC).
[0024] There are seven types of trichomes in tomato epidermis, including glandular and nonglandular trichomes. Among them, type I glandular hairs are the longest and visible to the naked eye. The leaves and stems of the Ln mutant LA3127 had a layer of dense epidermal hairs, the leaves appeared white, and the number of epidermal hairs increased significantly ( figure 1 ). In order to further determine which type of trichomes the Ln mutant has, we observed the mutant LA3127 and the control material Alisa Craig (AC) with a scanning electron microscope. Th...
Embodiment 2
[0030] Example 2 Development of Molecular Markers
[0031] According to the tomato classic map and high-density molecular genetic map, the Ln gene is located on the short arm of chromosome 3, and the corresponding segment of IL3-2 is between the two markers TG517 and TG366 ( image 3 ). In order to facilitate the development of molecular markers, we designed Indel molecular markers based on the differences in tomato Heizl1706 and LA1589 and S. pennellii reference genome sequences. According to the corresponding physical positions between TG517 and TG366, Indel markers in this interval were designed, and 10 pairs of molecular markers with stable polymorphisms between the two parents were screened out for Ln gene localization. The information of the localization markers is shown in Table 1 .
[0032] Table 1 Molecular marker information of Ln gene localization
[0033]
[0034]
Embodiment 3
[0035] Embodiment 3 Location of Ln gene
[0036] 116 normal trichomes were screened out from the F2 segregation population constructed by LA3127 and IL3-2. Using the Indel marker with stable polymorphism developed between TG517 and TG366 to conduct genotype analysis on 116 individual plants, the results showed that the Ln gene was located between the molecular markers LMP-4 and LMP-8 ( Figure 4 B), with 4 and 2 swaps respectively. According to the physical location of the two markers LMP-4 and LMP-8, the Ln gene is located in the range of about 2.35Mb between chromosome 3 2642776 and 5097282.
[0037]In view of the results of preliminary mapping, the Ln gene was located between molecular markers LMP-4 and LMP-8. Because the F2 segregation population constructed by LA3127 and IL3-2 received fewer seeds, we used the F2 segregation population constructed with LA3127 as the female parent and LA1589 as the male parent to further map the Ln gene.
[0038] 1243 individual plants ...
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