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Title:Genetics and expression of the anthocyanin biosynthetic pathway genes in soybean
Author(s):Wang, Chang-Sheng
Doctoral Committee Chair(s):Vodkin, Lila O.
Department / Program:Crop Sciences
Discipline:Crop Sciences
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:Ph.D.
Genre:Dissertation
Subject(s):Biology, Molecular
Abstract:The pigmentation of the seed coats in soybean is controlled by the genes I, R, and T. The results of the genetic study of the R mutable alleles, $R\sp*$, r-m55, and $r\sp*$ show that the dominance relationship between these alleles is changed from $R>r$-$m>r$ to r-m55 $>$ $R\sp*>r\sp*$; crossing between the mutable alleles generates high frequencies of both somatic and germinal mutations and results in switching the genotypes between the three alleles in this locus in all directions as $R\sp*\leftrightarrow r$-m55, $R\sp*\leftrightarrow r\sp*$, and r-m55 $\leftrightarrow\ r\sp*$. The mutable alleles have a tendency to stabilize in the F3.
A 200 bp soybean DFR gene fragment was cloned by polymerase chain reaction at 37$\sp\circ$C annealing temperature using two degenerate primers. The amino acid sequence was compared with the sequences of snapdragon, petunia, maize, and arabidopsis.
A phenol-LiCl based RNA extraction protocol was developed by the addition of the polyphenolic compound competitors, polyvinylpolypyrrolidone (PVPP) and bovine serum albumin (BSA), and the RNase inhibitors, heparin and proteinase K to extract RNA from the seed coats containing large amount of proanthrocyanidins. RNA extracted by this method is suitable for northern hybridization, reverse transcription, and polymerase chain reaction.
Expression of CHS mRNA was significantly reduced in the yellow seed coats (IRt); however, the DFR and PAL genes were expressed at the same levels as in the pigmented variety (iRt). Regulation of the pigmentation in soybean seed coats by the I alleles may be through the CHS gene in the anthocyanin biosynthetic pathway. Significant differences in CHS RFLP polymorphism were also found between I and i near isogenic lines digested with HindIII, SphI, SstI, and other enzymes. These data are consistent with two hypotheses--that I encodes a CHS gene or that I is linked to a CHS gene and regulates its expression in the seed coats.
Issue Date:1993
Type:Text
Language:English
URI:http://hdl.handle.net/2142/20556
Rights Information:Copyright 1993 Wang, Chang-Sheng
Date Available in IDEALS:2011-05-07
Identifier in Online Catalog:AAI9329193
OCLC Identifier:(UMI)AAI9329193


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