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Towel gourd fruit bitter taste gene LaBt and molecular marker used for auxiliary selection for bitter taste property of fruit

A fruit bitterness gene and molecular marker technology, applied in genetic engineering, plant genetic improvement, application and other directions, can solve the problems of time-consuming and laborious identification, inconsistent identification results, and inability to identify, so as to reduce identification costs, improve breeding efficiency, and save manpower physical effect

Active Publication Date: 2018-09-04
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 1) Identification is time-consuming and laborious;
[0007] 2) Different tasters have different sensitivities to bitterness; and the taste buds are prone to work fatigue, resulting in inconsistent identification results;
[0008] 3) It can only be identified at the fruit stage, not at the seed, seedling or plant stage, and plants bearing bitter fruits cannot be eliminated in advance;
[0009] 4) The bitterness of the fruit produced by the plant is easily affected by the environment, sometimes bitter and sometimes not bitter, resulting in inaccurate identification

Method used

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  • Towel gourd fruit bitter taste gene LaBt and molecular marker used for auxiliary selection for bitter taste property of fruit
  • Towel gourd fruit bitter taste gene LaBt and molecular marker used for auxiliary selection for bitter taste property of fruit
  • Towel gourd fruit bitter taste gene LaBt and molecular marker used for auxiliary selection for bitter taste property of fruit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Bitterness identification and genetic analysis of loofah fruit

[0038] Using multi-generation self-bred material 4-0-0-0 as the female parent (P 1 , not bitter, ribbed, hereinafter referred to as 4), 48-1-0-0 as the male parent (P 2 , not bitter, without ribs, hereinafter referred to as 48), crossed to obtain F 1 (4×48), and backcross (48×4)×48 to obtain backcross population BC1. to F 1 and backcross population BC 1 Identification of fruit bitterness phenotypes using sensory tasting methods: F 1 All of them exhibited bitter taste. Among the 87 backcross progeny materials, 41 were not bitter, 41 were bitter, and 5 were missing data due to too old and bitter taste of fruit or unhanging fruit (Table 1). The chi-square test shows that the actual ratio is in line with the theoretical ratio of 1:1, which proves that there is a single dominant bitter gene named LaBt in ribbed loofah.

[0039] Among them, the sensory tasting methods are:

[0040] When the frui...

Embodiment 2

[0045] Example 2 Utilize SRAP and SSR markers to construct loofah genetic map construction

[0046] 1. Primer screening

[0047] 192 pairs of SSR primers were selected from the primers developed by the inventor's research group and the primers of the previous research results (Wu et al., 2014) for PCR amplification on the parental materials '4' and '48', and 90 pairs were selected Co-dominant, stable amplification, and reproducible SSR primers are used for genetic mapping. 36 pairs of SRAP primer combinations with polymorphisms and clear bands were screened after amplification on parental materials with 324 pairs of SRAP primer combinations.

[0048] The loofah SSR amplification system is 10 μL, containing: DNA template 5ng / μL, forward and reverse primers 0.1 μmol / L each, 1×TaqBuffer (containing Mg 2+ ), dNTPs 0.1mmol / L. The reaction program was: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 50°C for 30 s, extension at 72°C for 1 min, 36 cy...

Embodiment 3

[0056] Example 3 Mapping of the Bitterness Gene LaBt in the Loofah with Ribs

[0057] 1. Preliminary mapping of the bitter taste gene LaBt in ribbed loofah fruit

[0058] The loofah fruit bitterness trait was used as a marker LaBt, and co-segregation analysis was carried out together with other markers, and the LaBt gene was located between the markers SGE292 and SGC196 on linkage group 6, with a distance of about 6.3 cM from SGE292 and a distance from SGC196 of 3.0cM.

[0059] 2. Fine mapping of the bitter taste gene LaBt in ribbed loofah fruit

[0060] Using bioinformatics analysis, the source sequences of the developmental markers SGE292 and SGC196 were anchored to the genome of cucumber through sequence comparison, and then the cucumber sequence between the two markers was downloaded from the cucumber genome website. After searching the data, it was found that the cucumber was in this region A bitterness-related gene Csa1G044890 was located, and the gene function was cyt...

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Abstract

The invention belongs to the field of molecular biology, and more specifically relates to a Luffa acutangula roxb bitter taste site LaBt genome sequence, a CDS sequence, a protein sequence, and a common towel gourd genome sequence corresponding to the site, a polymorphic marker for difference development according to a sequence between the above Luffa acutangula roxb and the common towel gourd. Though a molecular marker technology, a genetic linkage map of the towel gourd is constructed, the towel gourd bitter taste gene LaBt is positioned, an InDel molecular marker LuBt1-2 co-separated with the LaBt gene is obtained, an auxiliary selection system of the molecular marker is established, and the genome sequence of the site is obtained. The gene is helpful for auxiliary selection of the molecular marker for bitter taste property of towel gourd fruit, overcomes the disadvantages of sensory tasting and identification of the bitter taste of the fruit, and has the advantages of labor savingand time saving.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to the genome sequence of the cloned ribbed loofah fruit bitterness site LaBt, the genome sequence of the corresponding locus of common loofah, the SNP and InDel difference information between the two and its application, and the newly developed loofah. SSR markers and fine-mapping markers constructed on genetic maps, InDel molecular marker LuBt1-2 for identifying the bitterness traits of ribbed loofah, its primers and applications. Background technique [0002] Luffa is one of the important crops of Cucurbitaceae vegetables. There are two subspecies of loofah [Luffaacutangula (L.) Roxb.] and common loofah [Luffa cylindrica (L.) Rome.] commonly planted in my country, each with many excellent characters , if the loofah with ribs is crispy and not browned, the common loofah has high yield and strong resistance. Ribbed loofah and ordinary loofah are not bitter and can be eat...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/11C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/13
Inventor 汪国平秦永强吴宗斌崔丹丹汪文毅丁伟伦
Owner SOUTH CHINA AGRI UNIV
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