April - June 2007 - Kasetsart University
April - June 2007 - Kasetsart University
April - June 2007 - Kasetsart University
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<strong>Kasetsart</strong> J. (Nat. Sci.) 41 : 251 - 261 (<strong>2007</strong>)<br />
Anther Culture of BC 1F 1 (KDML105//IRBB5/KDML105) Hybrid to<br />
Produce Bacterial Blight Resistance Doubled Haploid Rice<br />
Supanyika Sengsai 1 , Surin Peyachoknagul 1 , Prapa Sripichitt 2 ,<br />
Amara Thongpan 1 and Pradit Pongtongkam 1 *<br />
ABSTRACT<br />
Maltose was found to be a better carbon source for callus induction in BC 1F 1 (KDML 105//<br />
IRBB5/KDML105) anther culture compared with sucrose. Statistical analysis, however, showed that<br />
increasing maltose or sucrose concentrations had no differential promotive effects on callus formation.<br />
One-step plantlet formation was found when maltose and NAA were supplemented together in the<br />
induction media. Adding 2 mg/l 2,4-D to the medium further increased the percentage of callusing<br />
anthers from 5.57% to 10.19%. However, the highest percentage of green plant regeneration was obtained<br />
(1.29%) from calli induced on N 6 medium without 2,4-D and subsequently cultured on regeneration<br />
medium containing MS supplemented with 2 mg/l BAP, 0.2 mg/l NAA, 300 mg/l casein hydrolysate,<br />
15% coconut water, and 30 g/l sucrose. AFLP analysis of all six anther-derived plants showed 57.3%<br />
to 67.12% recurrent parental alleles. After planting, seeds were detected in two out of six anther<br />
culture-derived plants indicating the occurrence of spontaneous chromosome doubling in these plants.<br />
Unfortunately, none of these six plants contained bacterial blight resistant gene (xa5) as detected by<br />
specific PCR-based RG556 marker and pathogen inoculation.<br />
Key words: KDML 05, anther culture, maltose, 2,4-D, AFLP, RG556, bacterial blight<br />
INTRODUCTION<br />
The production of haploid plants and<br />
doubled haploid plants from anther culture offers<br />
a rapid achievement of homozygous lines for early<br />
release of new crop varieties. Many desirable traits<br />
such as high grain weight, disease resistance, dwarf<br />
plant type and abiotic stress tolerance were<br />
introgressed into rice breeding population by<br />
culturing of anthers. Unfortunately, low<br />
percentages of both callus induction and plant<br />
regeneration are the principal constraints in<br />
establishing successful anther culture in some rice<br />
varieties especially in indica rice since these<br />
critical culturing responses are genotype<br />
dependent (Roy and Mandal, 2005). Consequently,<br />
the effective culture medium used for some rice<br />
varieties may not be appropriate for others, and<br />
the composition of culture media should be<br />
carefully selected when the anthers of particular<br />
rice variety was subjected to culture.<br />
Sucrose is generally added in rice anther<br />
culture media to serve as the standard carbon<br />
source and the osmotic regulator. However, many<br />
1 Department of Genetics, Faculty of Science, <strong>Kasetsart</strong> <strong>University</strong>, Bangkok 10900, Thailand.<br />
2 Department of Agronomy, Faculty of Agriculture, <strong>Kasetsart</strong> <strong>University</strong>, Bangkok 10900, Thailand.<br />
* Corresponding author, e-mail: fscipdp@ku.ac.th<br />
Received date : 19/07/06 Accepted date : 22/01/07