Genetics and expression of the anthocyanin biosynthetic pathway genes in soybean
Wang, Chang-Sheng
This item is only available for download by members of the University of Illinois community. Students, faculty, and staff at the U of I may log in with your NetID and password to view the item. If you are trying to access an Illinois-restricted dissertation or thesis, you can request a copy through your library's Inter-Library Loan office or purchase a copy directly from ProQuest.
Permalink
https://hdl.handle.net/2142/20556
Description
Title
Genetics and expression of the anthocyanin biosynthetic pathway genes in soybean
Author(s)
Wang, Chang-Sheng
Issue Date
1993
Doctoral Committee Chair(s)
Vodkin, Lila O.
Department of Study
Crop Sciences
Discipline
Crop Sciences
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Language
eng
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.
Use this login method if you
don't
have an
@illinois.edu
email address.
(Oops, I do have one)
IDEALS migrated to a new platform on June 23, 2022. If you created
your account prior to this date, you will have to reset your password
using the forgot-password link below.
