Detection and analysis method for heat resistance regulation and control capability of rhododendron heat shock transcription factor Hsfs
A technology of heat shock transcription factor and analysis method, which is applied in the field of detection and analysis of rhododendron heat shock transcription factor Hsfs to regulate heat tolerance, achieving short and fast results
Pending Publication Date: 2022-07-05
JIYANG COLLEGE OF ZHEJIANG A & F UNIV
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At present, most of the studies on heat tolerance and heat shock proteins and heat shock transcription factors are concentrated in plants such as Arabidopsis, tomato, sweet pepper and cucumber, and such research has not been widely carried out in Rhododendron
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[0032] In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the following are combined with specific embodiments and with reference to the accompanying drawings. Figure 1-12 , the present invention is described in further detail.
[0033] 1. Total RNA extraction and cDNA synthesis of Rhododendron
[0034] The leaves of Rhododendron with good growth and development were sampled, quick-frozen in liquid nitrogen after sampling, and then transferred from liquid nitrogen to -80°C ultra-low temperature freezer for preservation in preparation for the extraction of total RNA of Rhododendron.
[0035] 1. In this experiment, the RNA kit was used to extract the RNA of Rhododendron leaves. For the specific operation, please refer to the instructions of the RNA extraction kit of Tiangen Biochemical Technology Co., Ltd., and finally a high-concentration RNA solution was obtained.
[0036] 2. Use the kit of Tiangen Biochemical T...
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The invention belongs to the technical field of plant genetic engineering, and particularly relates to a method for detecting and analyzing the heat resistance regulation and control capability of a rhododendron heat shock transcription factor Hsfs. A target fragment is selected from full-length sequences of RpHsfC1 and RpHsfC1-like genes respectively, target gene fragments are amplified to be recombined with a virus vector, a VIGS vector is constructed, the VIGS vector is converted into rhododendron, and fluorescent quantitative expression analysis results show that the expression quantity of the target gene silenced by the VIGS is inhibited, which indicates that the target gene is successfully silenced, and the RpHsfC1-like gene can be used for preparing the RpHsfC1-like gene. A transgenic plant is obtained. Through analysis of photosynthetic fluorescence characteristics of the silent plant, it is found that target genes RpHsfC-1 like and RpHsfC-1 silenced by VIGS both have an influence on the heat resistance of rhododendron. According to the invention, a gene source is provided for cultivation of heat-resistant varieties of rhododendron by preliminarily identifying the heat-resistant regulation capability of the gene.
Description
technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a method for detecting and analyzing the heat-resistance regulation of Rhododendron heat shock transcription factor Hsfs. Background technique [0002] Rhododendron is a famous flower in my country, and it is also one of the top ten traditional flowers in my country. Rhododendron has a very wide geographical distribution. Rhododendron can grow in different vegetation zones from low altitude to high altitude, but Rhododendron is mainly distributed at altitudes of 1500-4000m. area, prefers a cool environment. At present, with the intensification of global warming, high temperature has become a global natural disaster, involving a wide range. However, temperature is an important factor affecting the growth and development of plants. Unsuitable growth temperature can easily affect the growth and metabolism of plants. Most of the excellent varieties of Rh...
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IPC IPC(8): C12N15/84C12N15/29C12Q1/6895C12N15/11
CPCC12N15/8218C12N15/8271C12N15/8205C07K14/415C12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/166
Inventor 金松恒申建双胡玥
Owner JIYANG COLLEGE OF ZHEJIANG A & F UNIV