Vector for editing tomato yellow leaf curl China virus (TYLCCNV) on basis of CRISPR/Cas9 system, and construction method and application of vector

Abstract

The invention discloses a vector for editing tomato yellow leaf curl China virus (TYLCCNV) on the basis of a CRISPR / Cas9 system, and a construction method and an application of the vector. The vectorcomprises pCambia 1300-BKG-g4, pCambia 1300-BKG-g5 and pCambia 1300-BKG-g6 which are constructed for a replication core region (an Iteron region) of the tomato yellow leaf curl China virus (TYLCCNV);and the vector also comprises pCambia 1300-BKG-g1, pCambia 1300-BKG-g2 and pCambia 1300-BKG-g3 which are constructed for the Iteron region of tomato yellow leaf curl China virus beta (TYLCCNB). The vector provided by the invention depends on a CRISPR / Cas9 system, can realize fixed-point editing of a TYLCCNV / TYLCCNB sequence, and has high editing efficiency.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for constructing a vector based on the CRISPR / Cas9 system target and editing the replication core region of Chinese tomato yellow leaf curl virus to inhibit geminivirus infection. Background technique [0002] Tomato yellow leaf curl China virus (Tomato yellow leaf curl China virus, TYLCCNV) belongs to Geminiviridae, Bean golden mosaic virus genus virus. The virus is often accompanied by the satellite molecule Tomato yellow leaf curl China betasatellite (TYLCCNB) in the field. The disease complex formed after the combined infection of TYLCCNV and TYLCCNB has caused destruction to vegetable crops such as tomatoes in the field in my country. sexual hazards. The virus is mainly transmitted by Bemisia tabaci, and the infected tomato plants grow slowly or stagnate, with serious dwarfing, new leaves chlorotic, yellow, smaller, and curled at the edges. At present, ...

AI Technical Summary

Problems solved by technology

Previously, by using Cas9 technology to target the IR region of other geminiviruses, the disease resistance of plants obtained was not satisfactory

Under the current conditions, there are still many constraints in the prevention and control of geminiviruses in the field: (1) plant viruses are known as "plant cancers", and field control is difficult. Due to the unsatisfactory biological, physical and chemical control effects, there are currently many anti-virus projects in the field. Focus on prevention and combine prevention and control; (2) Geminiviruses mostly rely on insect vectors for transmission, so controlling crops in the field to eat insects is the most effective way to cut off its transmission route, so that large-scale use of chemical reagents cannot be avoided, and pesticide residues Problems and non-target biosafety issues are in front of people, and it is difficult to solve in a short period of time; (3) In terms of breeding, the breeding of geminivirus-resistant crop varieties is slow, and the field effect is not ideal, and there are potentially great uncertain factors

(4) At present, the use of CRISPR / Cas9 technology to edit plant virus sequences mostly selects in the IR region of geminiviruses, and the antiviral effects vary greatly among different viruses, and there is no good specificity

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