Method for acquiring disease-resistance expression of agrobacterium-mediated potato transgenic hrap

Abstract

The present invention discloses a method for attaining potato plates which are disease resistant by utilizing agrobacterium-mediated hrap gene. After the combined inoculation of the method and the bacteria allergic protein harpinPss, the allergic reaction caused by harpinPss can be promoted in the intercellular space of all inoculated plates, and the polymeride of harpinPss is dissociated, so the disease resisting effect is better, and the disease resistance of the potato plates can be promoted. The hrap gene is introduced in the potato plates through the agrobacterium-mediated, thus resulting that the potato plates have the ability to resist the late blight and the bacterial wilt. The present invention has the technical proposal that the pre-cultured explants are inoculated in the agrobacterium germ liquid to be soaked, explants after being soaked select the tube seedling which can root after two times of root selections by co-culturing, callus of induce, bud inducting and rootage cultivating and selecting, and the potted transformed plants are transferred after the mass propagation, the transformed plants select positive plants by the methods of the PCR detection and the Southern hybridization detection, and then the potato plates which are disease resistant can be obtained by the in-vivo inoculation resistance identification. The present invention improves the callus of inducing and the bud inducing of the explants, and enhances the agrobacterium-mediated ability. That the roots of the explants are selected by two times can eliminate the false positive plates in the transformation process , avoid the continuing cultivation to the false positive plates, save the labour force and decrease the manufacturing cost.

Description

technical field [0001] The invention relates to a method for using agrobacterium to mediate the introduction of hrap gene in potato to obtain the expression of disease resistance. Background technique [0002] Potato is the fourth most important food crop in the world, and it is also a vegetable, feed, processing and industrial raw material crop. Now there are 148 countries that grow potatoes in the world, with a total output of 300 million tons. Research by the International Potato Center (CIP) shows that worldwide demand for potatoes will be expected to increase by 20% by 2020, surpassing the growth of rice, wheat, and corn; times. [0003] Our country has unique advantages in producing potatoes in terms of geographical and climatic conditions. The perennial planting area of ​​potatoes in my country is about 4.7 million ha 2 , with a total output of more than 70 million tons. The output and area account for one-fifth and one-fourth of the world's total respectively, r...

AI Technical Summary

Problems solved by technology

However, potato is an autotetraploid crop, and it is difficult to directly use it due to reproductive barriers, long breeding cycle, and complicated process to improve varieties by conventional breeding methods.

At the same time, the vertical resistance gene of the wild species is easy to lose resistance due to the rapid variation of the pathogenic race; the horizontal resistance gene alone is not enough to effectively resist the invasion of late blight, and is easily affected by environmental conditions, resulting in the pandemic of late blight

Potatoes suffer from reduced quality and poor potato shape after being infected with the disease, resulting in a low rate of commercial potatoes, which is far from meeting the needs of processing and the market. At the same time, it affects economic benefits and the enthusiasm of potato farmers, and restricts the production and processing capabilities.

But there is no report on transformation and disease resistance expression of hrap gene in potato

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