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Application of related gene for improving content of mogroside

A content and gene technology, applied in the field of plant biology, can solve the problem of incompletely clear biosynthetic pathway of mogroside

Inactive Publication Date: 2019-04-26
怀化兴科创生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biosynthetic pathway of mogroside in Luo Han Guo is not fully understood

Method used

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  • Application of related gene for improving content of mogroside
  • Application of related gene for improving content of mogroside
  • Application of related gene for improving content of mogroside

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1, Three Generation Sequencing of Luo Han Guo Whole Genome

[0054] The seedlings of Luo Han Guo samples were purchased from Yongfu District, Guilin City, Guangxi Province, which is the most widely planted green fruit variety. The seedlings were planted in Zhongfang County, Huaihua City, Hunan Province (such as figure 2 shown). The roots, buds, leaves, male and female flowers, and fruits of different periods after pollination were collected from August to October. The samples were frozen in liquid nitrogen and stored in a -80°C freezer. The stem tip of Luo Han Guo was used for genome sequencing experiment due to the extraction of genomic DNA.

[0055] In this study, high-quality DNA was extracted, and two PacBio RS II libraries with an insert fragment of 20kb were constructed. The third-generation sequencing platform PacBio RS II was used to perform non-amplified long-fragment sequencing of the DNA. A total of 44 SMRT cells were obtained to obtain Luo Han Gu...

Embodiment 2

[0059] Example 2. Sequencing, assembly and analysis of Luo Han Guo transcriptome

[0060] The varieties sequenced by the transcriptome of Luo Han Guo were consistent with those sequenced by the genome (such as Figure 5 Shown), are all green fruits, introduced from Yongfu County, Guangxi Province, and planted in Zhongfang County, Huaihua City, Hunan Province. At different growth and development stages, 6 leaves of female plants, leaves of male plants, leaves of adjacent fruits, fresh young roots, fruits three days after fertilization (3DAA), and fruits 20 days after fertilization (20DAA) were collected. For each sample, there are at least two biological replicates. After all samples are collected, use ddH 2 O was washed three times, then dried with absorbent paper and quickly wrapped and stored in liquid nitrogen for later use.

[0061] RNA extraction was carried out according to the steps indicated in the kit. After the samples were tested to be qualified, 3 μg of total RN...

Embodiment 3

[0063] Embodiment 3, the cloning of Momordica grosvenori epoxide hydrolase gene (Sgeph7)

[0064] Whole-genome sequencing and transcriptome sequencing were used to obtain the sequence information of the epoxide hydrolase gene (Sgeph7) of Luo Han Guo, respectively. The genomic DNA and RNA of the mature fruit of Luo Han Guo were extracted, primers were synthesized by PCR technology, and the gene fragment, cDNA fragment and promoter were cloned, and the gene was named Sgeph7. It includes promoter DNA fragment (SEQ ID NO:4); cDNA sequence (SEQ ID NO:2), genome sequence (SEQ ID NO:1) and amino acid sequence (SEQ ID NO:3).

[0065] Figure 7 It is the evolutionary comparison of the epoxide hydrolase of Luo Han Guo and Cucurbitaceae plants cucumber, zucchini and mountain gourd, in which green is cucumber; red is zucchini; yellow is mountain gourd; blue is Luo Han Guo. The genetic relationship between the EPH gene of Luo Han Guo and the EPH genes of Cucurbitaceae plants such as cucu...

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Abstract

The invention discloses a mogroside synthesis regulation gene Sgesph7 and application thereof, and belongs to the field of biotechnology. According to the mogroside synthesis regulation gene Sgesph7 and the application thereof, by means of whole genome sequencing of Siraitia grosvenorii(Swingle)C.Jeffrey and the analysis of the RNA-seq sequence of different tissue, secondary metabolic pathways associated with the tissue specificity and characters are utilized, and therefore the Siraitia grosvenorii(Swingle)C.Jeffrey epoxide hydrolases Sgeph7 gene with Siraitia grosvenorii(Swingle)C.Jeffrey fruits specifically expressed; and the content of the mogroside in the Siraitia grosvenorii(Swingle)C.Jeffrey fruits can be regulated and controlled by adjusting the expression of the gene, and the geneis used for obtaining a new variety or new strain of the Siraitia grosvenorii(Swingle)C.Jeffrey with the high mogroside content and has important theoretical and practical significance in research ofthe taste mechanism of fruits of cucurbitaceae plants and the breeding work of the taste of the fruits of the cucurbitaceae plants.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and specifically relates to a method for breeding Cucurbitaceae plants, including increasing the content of sweet glycosides and improving the taste, and more specifically relates to a nucleic acid molecule of the regulating gene Sgeph7 for sweet glycosides content and its molecular breeding material and its use in Cucurbitaceae. Application in fruit improvement and breeding work. technical background [0002] Luo Han Guo (Siraitia grosvenorii (Swingle) C. Jeffrey) has been used as traditional Chinese medicine for centuries. Traditional Chinese medicine uses it to treat hypertension, tuberculosis, gastritis, asthma, pertussis, acute and chronic bronchitis and acute and chronic tonsillitis, etc. In recent years, studies have also found that Luo Han Guo contains anti-cancer ingredients, which has attracted great attention from the medical circles at home and abroad. Luo Han Guo is rich in activ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/55A01H5/00A01H6/00A01H6/34
CPCC12N9/14C12N15/8245C12Y303/02003
Inventor 夏勉韩雪何航邓兴旺
Owner 怀化兴科创生物技术有限公司
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