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Inheritance of a No Vine-type Cucurbita Moschata and Establishment of Its High Frequency Regeneration System

Author: ZhangYaFeng
Tutor: CaoJiaShu
School: Zhejiang University
Course: Olericulture
Keywords: Petiole cotyledons ELISA Endogenous hormones Biological traits No Man Genetic Analysis Regeneration Pumpkin
CLC: S642.1
Type: Master's thesis
Year: 2007
Downloads: 114
Quote: 1
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Abstract


Pumpkin (Cucurbita moschata (Duoh.ex Lam.) Duch.cx Poiret) for the Chinese Pumpkin species Cucurbitaceae Cucurbita. Currently, most of cultivation of pumpkin long vine varieties, no vine or small the short vine dwarf varieties. The small pumpkin without vine varieties cultivated in the required space, easier to manage, suitable for intensive cultivation, a great advantage in the production. Thus, no vine breeding traits for pumpkin significance. Cucurbita worldwide plant viral disease serious harm, conventional breeding proportion of its resistance breeding is still great, but due to the long life cycle, genetic traits instability, growing attempts by means of genetically modified new varieties. The corresponding virus coat protein gene into a pumpkin species in vivo, can greatly increase the resistance of plants to improve yield and quality. Pumpkin No Man mutations in body, scandent related genes can also be transferred to the plants in vivo to study the control of the mechanisms of the melons plant vine elongation. The success of transgenic technology needs to be built on the basis of efficient regeneration system. The study of the pumpkin 'No Man' selfed offspring found no vine traits homozygous plants Vine and biological characteristics of the observation and analysis of this study to control the genetic background of the pumpkin vine trait gene. Build pumpkin 'No Man No. 1' high frequency regeneration system based on the enzyme-linked immunosorbent assay (Enzyme-linked Immunosorbent Assays, ELISA) determination of endogenous hormones in different the seedling age Daibing cotyledons explants content. To analyze the relationship between endogenous hormone content and proportion of the initial explants and explant regeneration frequency. Currently get the following, some preliminary results: (1) on the pumpkin 'No Man No. 1' selfing offspring found no vine traits homozygous plants for No Man parent, long vine plants as long tendril parent. No Man parents, long vine parent, the pros and cons of hybrid generation (first filial generations, F 1 ), the reciprocal backcross generation (first back crosses, BC 1 s ) and hybridization second generation (second filial generation the F 2 ) plants Vine observation results show that, no vine with long vine traits can distinguish three true leaves period, no vine pro this all for no Man traits, long vine pro this all performance long vine traits the reciprocal hybrids F 1 s offspring is all no vine plants reciprocal backcross BC 1 s exhibit Man and the long vine plants 1:1 segregation ratio of F 2 no vine and long vine plantations segregation ratios of 3:1 progenies. This indicates that the the the Pumpkin Vine is controlled by a pair of alleles, control vine traits dominant gene on the long tendril trait gene expression. No vine traits produce dominant mutation is recessive gene control long vine traits. No Man parents long vine parent biological traits observed results, the long vine plants the male flowers was much more than the profusion of plants, flowering vine plants earlier than long vine plants 8d. No significant difference in the long vine plants vine plants leaf and petiole size. No Man pro-this and F 1 plants in the observed biological traits were not significantly different. (2) through the study seedling age, hormone concentration and type of explants body sources of renewable process, the establishment of a pumpkin 'No Man No. 1' high frequency regeneration system. The findings of the half cotyledon, Daibing cotyledons, petioles, root and leaf block five sources explant regeneration effect explants efficiently induce callus, but only Petiole cotyledons adventitious bud differentiation and adventitious buds is not generated by the callus, but directly from the cotyledon base. This adventitious buds from our research is the direct organogenesis pathway, the cotyledons base directly to key parts of the differentiation of adventitious buds. The results show that using different concentrations of BAP induced adventitious buds in four concentration gradient (for 0, 0.5 mg · L , -1 1.0mg · L -1 , 2.0mg · L -1 ), when the BAP does not exist no adventitious buds generated three other concentration does not have a significant effect on the regeneration frequency, BAP is necessary for the adventitious buds induced but its concentration on regeneration frequency is not significantly affected. The research results show that 4 ~ 7d the seedling age Daibing cotyledon explant regeneration process, BAP when used alone, showed a regular change in frequency of regeneration and explant regeneration frequency: 4 to 5d regeneration frequency declined after rising and increased to a maximum at 7d. (3) of Different Ages Daibing the cotyledon explants of endogenous hormone test results show that the highest content of ZR and IAA 4d seedling age, respectively 0.41 ± 0.069nmol, · g -1 FW -1 FW, after With the increase in seedling age decreased gradually to 7d at their lowest, respectively, 0.17 ± 0.037nmol · g -1 and 0.96 ± 0.223nmol · g FW 0.40 ± 0.152nmol · g -1 FW, and the relationship between the two and regeneration frequency wireless. of iPA and IAA / (IPA Zr) change and regeneration frequency showing a linear relationship. Petiole Explants of Cotyledon highest regeneration frequency 7d seedling age, while endogenous iPA the content and IAA / (iPA in ZR) values ??have the highest; 5d seedling age, the lowest frequency of regeneration of cotyledon explants, the corresponding inner The lowest values ??of the source iPA content and IAA / (iPA ZR). The endogenous GA 3 measurement results show that high levels may reduce the frequency of explant regeneration. The above results show that, in this experiment the pumpkin vine traits dwarf mutant is generated because the the control long tendril traits recessive gene dominant mutation. Build pumpkin 'No Man No. 1' Regeneration System of research show that the best for 7d the seedling age Daibing cotyledon explants of different concentrations of BAP on regeneration is not significant. The endogenous hormones measured results show that the relationship between Petiole cotyledon explants seedling age and frequency of regeneration, may be due to the result of changes in content and proportion of endogenous hormones.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Melons > Pumpkin
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