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The Effect of Sealing Materials on Immature Wheat Embryos Culture, and the Cloning and Analysis of Alpha-tubulin Genes in Regeneration Organs

Author: YuXueJun
Tutor: WuBiHua
School: Sichuan Agricultural University
Course: Crop Genetics and Breeding
Keywords: Wheat Immature embryo Tissue culture Regenerated flower buds Renewable root Young panicle α - tubulin cDNA Amino acids
CLC: S512.1
Type: Master's thesis
Year: 2010
Downloads: 29
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Abstract

YA-1 immature embryo of the wheat varieties as explants explore the effects of sealing material Immature Embryo direct regeneration, and the the immature embryos organization on flower buds reproductive organs and adventitious roots vegetative organs α-tubulin protein gene structure-specific research, the main results are as follows: 1. silicone plugs, YA-1 immature embryo callus redifferentiation the tampon two different sealing materials redifferentiation culture conditions the adventitious buds rates, adventitious roots rate browning rate is extremely significant differences (P lt; 0.01). The former is only 19.21% of the rate of regeneration of adventitious buds, which is up to 44.86%; the adventitious roots rate of the former was significantly lower than the latter (19.69% lt; 39.90%); browning former was significantly higher than the latter (49.62% gt; 8.37 %). The YA-1 immature embryo callus regeneration from wheat varieties flower buds and regeneration of root organs, as well as field normal growth and development of young spike 1A chromosome amplified cDNA open reading frame (ORF) F-1A, R-1A the length of the S-1A are 1353 bp, the sequence similarity of 100%. The length of the deduced amino acid sequence contains 450 amino acid residues based on their nucleotide sequences, the theoretical molecular weight are the 50 kD, isoelectric point are 4.71. In its amino acid sequence 142 to 148, there is a microtubule protein signal fragment GGGTGSG, the C-terminal has a tyrosine residue, the N-terminal are with MREII sequence. F-1A, R-1A, S-1A with the published T. aestivum'Quantum 'aboveground vegetative organs of wheat-DQ435673 the cDNA of the α-tubulin gene sequence similarity of 99.41%, with which there are 32-bit point nucleotide differences. CDNA open reading frame (ORF) was amplified from the above materials 3B chromosome obtained F-3B, R-3B, the length of the S-3B are 1356 bp. Based on their nucleotide sequences, the length of the deduced amino acid sequence contains 451 amino acid residues, the theoretical molecular weight are 50KD. F-3B, S-3B isoelectric point is 4.65, and the R-3B and the isoelectric point was 4.70. In their sequence of amino acids 142 to 148 there is a fragment GGGTGSG microtubule protein signal, the C-terminal has a tyrosine residue, the N-terminal are with MRECI sequence. F-3B and S-3B are reported T. aestivum'Quantum 'aboveground vegetative organs of wheat α-tubulin gene DQ435662 sequence similarity up to 100%. R-3B from the YA-1 immature embryo callus regeneration adventitious roots and F-3B, S-3B and DQ435662 on 529 points is replaced by a G to A nucleotide differences. 1A and 3B on the chromosome of the α-tubulin gene of the nucleotide sequence between the similarity due to the nature of the organ donor materials vary. Which flower buds reproductive organs, whether it is from the regeneration cultured flower buds, or the the field normal growth and development of the young spike, its cDNA sequence similarity are 84.69%, 80.68% and adventitious root similarity cloned from vegetative organs. Three materials tested in the amino acid sequence of the α-tubulin 1A and 3B chromosome similarity are 91.8%, their differences were seen in the 439 peptides. Thus, the α-tubulin protein wheat 1A and 3B chromosome genes and the amino acid sequence structure of both the homogeneity and heterogeneity exists, and in the tissues and organs of the different nature of the two chromosomes measured on α - tubulin gene and amino acid sequence structure with a high degree of stability, α-tubulin gene is highly conserved characteristics. 3 Comparative Analysis of the F-1A from wheat and reported maize, rice, soybean, cotton, Arabidopsis, zebrafish, found after the human amino acid sequence of the α-tubulin gene, the F-1A and reference 7 strip up to 93% sequence homology. Homologous tree analysis, F-1A with Gramineae corn, rice sequence together, which confirms its relatively close genetic relationship on the level of α-tubulin.

The Effect of Sealing Materials on Immature Wheat Embryos Culture, and the Cloning and Analysis of Alpha-tubulin Genes in Regeneration Organs

Abstract

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