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Stress
Tolerance in Soybeans
E.
Lewis Myles and Deborah Long
Cooperative
Agricultural Research Program
Seminar Series
Tennessee State University, Nashville, TN
February 10,
1999
Introduction. Conventional plant breeding
techniques for the identification of genes take a very long time and do
not have the versatility of biotechnology or molecular procedures. If a
gene sequence is identified, it can be used in a wide variety of ways.
Objectives. To screen selected soybean cultivars
for metal, drought, and disease tolerance and to use molecular
techniques for the identification of genes.
Methodology. A 10-day screening technique was
used to identify tolerant and sensitive cultivars. This procedure
consisted of 60 seeds per experiment and was repeated three times. Each
experimental group had 20 seeds in 10 sterile tubes. The average of each
tube was analyzed in Excel Analysis of Variance (ANOVA). Total protein
was extracted from hypocotyls and roots. These proteins were analyzed by
one- and two-dimensional gel electrophoresis. Total RNA was extracted
from which messenger RNA was isolated. cDNA was synthesized and analyzed
using random probes.
Results. Tolerant and sensitive cultivars were
identified for metal (aluminum and cadmium), drought and disease.
Differential display was used to identify metal tolerant genetic
sequences.
Impact. Two graduate and five undergraduate
students were trained in tissue culture and plant molecular techniques
for the fiscal year 1998. Both graduate students are working in
molecular research, and three of the four undergraduates are now in
Graduate School. At least 10 to 15 of the nucleotides were identified
that could lead to the identification of genes in eukaryotic systems.
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