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Studies on Agrobacterium-Mediated Transformation of Apple Rootstock M.26 with MdNHX1 Gene

Author: LiYongHong
Tutor: MaFengWang
School: Northwest University of Science and Technology
Course: Pomology
Keywords: Apple rootstock 'M.26' MdNHX1 gene Agrobacterium-mediated Salt stress
CLC: S661.1
Type: Master's thesis
Year: 2010
Downloads: 104
Quote: 1
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Abstract


Soil salinity stress seriously affect China's agricultural production and, more recently, by the constitutive expression of plant Na / H antiporter gene, a variety of genetically modified plants significantly improved salt tolerance, that cultivate salt-tolerant crops varieties through this strategy has broad prospects . So far, for fruit tree plants NHX gene function and gene transfer NHX Fruit Trees still small. In this trial, the apple rootstock 'M.26' plantlets blade for the material, the use of Agrobacterium tumefaciens-mediated method, from the salt tolerance the apple rootstock Luo-2 (Malus domestica Borkh.) Clone of Na / H antiporter gene (MdNHX1) apple rootstock 'M.26' translational research. Some physiological indicators simultaneously genetically MdNHX1 gene apple rootstock 'M.26' exogenous gene expression characteristics and integration and salt stress conditions were studies in order to take advantage of MdNHX1 genetic transformation of fruit trees, salt-tolerant transgenic plants to try. The main contents are as follows: 1 apple rootstock 'M.26' plantlets leaves receptor apple rootstock 'M.26' adventitious bud regeneration effects of different hormone concentration ratio, optimize the apple rootstock ' M.26 'Leaf Regeneration System. The results show that the induction of apple rootstock 'M.26' best medium for adventitious bud regeneration MS 4.0mg / L 6-BA 0.2mg / L NAA 30g / L sucrose 7.0g / L agar (pH5.8), renewable rate of 96%. By Agrobacterium tumefaciens-mediated method, leaves the receptor the transformation apple rootstock 'M.26 MdNHX1' 15 kanamycin resistance strains. Resistant lines obtained PCR test result, 11 lines can be detected to the target gene (MdNHX1) and resistance selection gene (NPT II). To 1WNHX1 and npt Ⅱ probe-positive strains were detected by PCR in 11 Southern blot analysis, the results showed that 6 positive strains, respectively named N2, N3, N4, N7, N8 and N9. Positive strains Southern blot analysis of the target gene RT-PCR assay results were positive. 6 lines 4 Real Time RT-PCR technology to study the different transgenic lines MdNHX1 gene expression levels. The results show that the gene expression levels of different transgenic lines MdNHX1 differences. Transgenic lines both showed higher expression levels than the control plants, which the N7 strains showed the highest and most stable MdNHX1 gene expression levels. 5. Analysis of the changes in some physiological indices of different transgenic lines in different concentrations of NaCl stress conditions. The results show that MdNHX1 gene expression, in varying degrees, to improve the roots of transgenic plants under salt stress of Na, K content; while reducing the Na content of the leaves that can improve the salt tolerance of plants through transgenic.

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CLC: > Agricultural Sciences > Gardening > Fruit trees gardening > Pome > Apple
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