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Breeding method capable of improving content of potassium in tobacco leaves

A technology for tobacco leaves and potassium content, which is applied to other methods of inserting foreign genetic materials, botanical equipment and methods, angiosperms/flowering plants, etc., and can solve the problems of small descendant groups, long segregation generations, genetic instability, etc. To achieve the effect of easy stability, simple and convenient operation, and avoid comprehensive reorganization

Inactive Publication Date: 2012-11-21
HUNAN AGRICULTURAL UNIV
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  • Description
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Problems solved by technology

[0003] However, conventional breeding has problems such as incompatibility of distant hybrids, unstable hybrid traits, and long segregation generations; although somatic cell hybridization technology has overcome the phenomenon of hybrid incompatibility to a certain extent, it has problems such as low regeneration frequency, small population of mutant offspring, Problems such as somaclonal variation and genetic instability; since Hess proposed that pollen grains can absorb exogenous DNA, total DNA introduction technology has been widely used in crop breeding, which overcomes the reproductive barriers in conventional breeding and makes up for In the current situation where the genetic background of tobacco is still unclear, the source of good genes is limited, and transgenic research is still negatively affected by trade barriers, it is a practical and feasible method. A shortcut to breed improvement

Method used

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  • Breeding method capable of improving content of potassium in tobacco leaves
  • Breeding method capable of improving content of potassium in tobacco leaves
  • Breeding method capable of improving content of potassium in tobacco leaves

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Experimental program
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Embodiment 1

[0015] Embodiment 1: the performance of main characters of tobacco progeny after the method of the present invention is implemented

[0016] Follow the steps below for DNA introduction:

[0017] (1) Extraction of donor DNA: Take 0.2g of young tobacco leaves and put them into an inkstone bowl → add 700μl of 2×CTAB extract solution for grinding and then transfer to a 1.5ml centrifuge tube → incubate at 65°C for 20-30min → add after cooling Volume of chloroform: mix well with isoamyl alcohol for 10 minutes→centrifuge at 10000r / min for 10 minutes→take supernatant to a new tube→add 100μl 5M NaCl and 1ml ice-cold ethanol to precipitate DNA→pick out DNA precipitate, wash and dry with 70% ethanol→dissolve In 100 μl TE; the formula of the 2×CTAB extract is: 2% CTAB, 1.4M NaCl, 100 mM Tris-HCl, 100 mM EDTA, pH8.0, 2% β-mercaptoethanol; the chloroform: isoamyl alcohol The volume ratio is 24:1;

[0018] (2) Import method: Select a flower that grows vigorously and has opened in the middl...

Embodiment 2

[0032] Embodiment 2: the performance of tobacco progeny tobacco leaf quality after the method of the present invention is implemented

[0033] 1. Tobacco leaf appearance quality performance

[0034] The evaluation results of tobacco leaf appearance quality showed (Table 4): Honghua Dajinyuan and D 9 -05 and D 9 The performance of -12 strains was basically the same in terms of maturity, while the D 9 The -06 strain matures late; the most oily is Honghua Dajinyuan, followed by D 9 -06 strain; Honghua Dajinyuan and D 9 -05, D 9 -06 and D 9 Most of -12 structures are loose; D 9 -05, D 9 -06 and D 9 -12 is better than K327 in fullness, uniformity and color. This shows that the appearance quality of the new line is slightly better than that of the control Honghua Dajinyuan, but the difference is not significant.

[0035] Table 4 Comparison of appearance quality evaluation of different lines of tobacco leaves

[0036]

[0037] Note: B, C, and X in the position represent t...

Embodiment 3

[0048] Embodiment 3: the expression of potassium-related genes in tobacco progeny after the implementation of the method of the present invention

[0049] Real-time fluorescent quantitative PCR method was used to measure the changes of mRNA transcription levels of related high-potassium genes in different tobacco high-potassium new lines and controls ( figure 1 ). The results showed that: in different new strains, compared with the control material, D 9 -05, D 9 -06 and D 9 The expression of high-potassium genes in the -12 new line is the most obvious, which shows that the new line obtained in this experiment has a high potassium content; on this basis, among the different potassium-related genes, Ntpk1 gene, Cbl9 gene and Gork gene are in the The expression levels in the three new lines were relatively high; the Ntkc1 gene in D 9 The expression level in the -05 new line was high, but in D 9 -06 and D 9 -12 new lines are relatively low; Tork1 gene in D 9 The expression ...

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Abstract

The invention provides a breeding method capable of improving the content of potassium in tobacco leaves. According to the invention, total DNA of the high potassium plant pokeweed is introduced into Honghuadajinyuan, a cultivated variety of tobacco, by using a pollen tube pathway method, which results in substantial increase of the content of potassium in later-generation tobacco leaves of Honghuadajinyuan, and therefore, a rich source of variation is provided for breeding and improvement of new varieties of tobacco and innovation of germplasm resources.

Description

technical field [0001] The invention relates to a breeding method for increasing the potassium content of tobacco leaves. Background technique [0002] According to a large number of statistics, potassium is the nutrient element with the highest absorption and body content in flue-cured tobacco, and it is also an important limiting factor affecting the further improvement of flue-cured tobacco quality in my country. However, most tobacco planting soils in my country cannot provide enough potassium to supply flue-cured tobacco to meet the needs of high-quality tobacco production. Therefore, a large amount of potassium fertilizer needs to be invested in production. The continuous application of large amounts of chemical fertilizers not only increases the production cost of tobacco farmers, but also accelerates the deterioration of soil physical and chemical properties. Unfortunately, the average potassium content of high-quality tobacco leaves in my country is still low, at ...

Claims

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Application Information

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IPC IPC(8): C12N15/87A01H5/00
Inventor 戴林建钟军徐双红王翔朱列书符建国夏凯
Owner HUNAN AGRICULTURAL UNIV
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