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aqueous solution mixed by the 0.05% colchicine and
2% DMSO to intermittently treat Baofeng 2 pea seeds
for 48 hours, and fertile autotetraploid was found by
morphological and cytological identification and com-
parison on the latter generations of treated pea. Signifi-
cant differences of plant shape between autotetraploid
and its diploid were shown as below: high plants , thick-
ened stems and tendrils, total leaf area per plant being
larger, leaf color greener, flowering stage delayed, bigger
pollen grains and pollen mother cell, larger pod , self-fer-
tility, and bigger seed. In this research, 71.86% mutants
were obtained by treatment of 0.4% colchicine, the
greater the treatment concentration was, the higher the
variation rate would be, and the more abundant the
variation types would be. However, the mutants of
better yield factors was obtained in the treatment of
0.2% colchicine.
The method of 0.2 and 0.4% colchicine treating for 12
h is effective for inducing mutation of azuki bean,
with fast homozygosity, abundant variation, but chrom-
some doubling was induced. These mutants aslo provi-
ded valuable materials for the breeding of azuki bean
and studies on gene mapping and positional cloning.
3 Materials and methods
3.1 Materials and colchicine treatment
Jingnong 6 (JN6) variety of adzuki bean was provided
by Azuki Bean Research Group of Beijing University
of Agricultural. 1 000 seeds of JN6 were treated with
0.2% and 0.4% colchicines for 12 hours, respectively.
The treated seeds were planted in experimental plots
of Beijing University of Agricultural with 40 cm row
spacing, 15 cm plant spacing and double-grain dibble
seeding from 2006 to 2008.
3.2 Trait investigation and data analysis
During the growing period, seedling stage, trefoil stage,
flowering stage, pod-setting stage and maturing stage
were investigated, and seedling rate and plant survival
rate were counted. All plants were harvested when mat-
ured and dried for plant laboratory tests with items inclu-
ding plant height, start nod of branches, branch number,
pod length, pod width, pod number per plant, the number
of seeds per plant, 100
-
seed weight and yield per plant.
All data were processed by SPSS statistical software.
3.3 Seed Color Mensuration
Seed color was tested with CR
-
200b Color Difference
Meter, manufactured by Japan-based Minolta Camera
Co., Ltd., which used the O/D mode of diffused illumi-
nation and vertical light interception. L* indicated bri-
ghtness index, and the greater the value was, the brighter
it would be; a* and b* were nominated as redness and
yellowness index, indicating the red and yellow degree
of azuki bean seeds respectively. The greater a* value
was, the redder the color would be; the greater b* value
was, the fresher the color would be. 5 plump seeds were
selected from each seed color mutant and control, for
measuring 3 times, and counted the average.
Authors’ contributions
Baomei Wu carried out this research, analyzed data and drafted partial
manuscript. Hongxia Liu and Bo Zhao, Xing Tong helped with the analyses
and wrote substantials parts of the paper. Xing Tong and Qingmu Zhao
worked on obtaining data. Ping Wan conceived the project, performed the
experiment designs, super- vised the data analysis and revised this paper. All
authors read and approved the final manuscript.
Acknowledgements
This study was jointly funded by Science and Technology Research Fund of
Educational Committee of Beijing City (KM201010020004), Blossom Edu-
cation Through Talent Introduction Project (PXM2007
-
014207
-
04453) and
Top-notch Talent Project (PXM2007
-
014207
-
021717) of Beijing Muni-
cipal Education Commission. We thank Guangjun Lei (Azuki Bean Research
Group, College of Plant Science and Technology, Bejing University of
Agriculture) for material planting and the experiment. This paper is for com-
memorating respectful professor Wenlin Jin.
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