1. Improving on TDK1-Aroma by Marker Assisted Selection (MAS)
Seminar on 15thNAFRI Anniversary Symposium on Agriculture and Forestry Research for Development
April 9th-10th, 2014 at NAFRI, Vientiane, Lao PDR
By Chanthakhone Boualaphanh, Kongpanh Kanyavong, Souvanh Thadavong and staffs
Agricultural Research Center (ARC), Lao PDR

2. Aromatic rice
Highly prized by most rice consumers, and many countries cultivate traditional and improved aromatic varieties.
2-acetyl-1-pyrroline (2AP) is the major aromatic compound in rice, and several studies show about 100 other volatile compounds in rice
(Buttery et al., 1983; Lorieux et al., 1996)

3. Aromatic traditional Lao rice varieties
There are many aromatic rice
All traditional varieties
Low yielding
Growing in only wet season
such as: Hom nang nuane, Kai noy, Homthong ……..
Hom nang nuane
Kay noi

4. Achievements of Lao Rice Research Program
There are many vars. Are released:
TDK 1 - TDK 13
TSN 1 – TSN 9
PNG 1 – PNG 6
All vars. are non aromatic rice
Before, for developing vars. we use only conventional breeding

5. Technique to select aroma plant
Sensory test
Check 2AP by using gas chomotograph
Marker Assiated Selection (MAS)

6. Rice Research Program
In 2004, Laos (ARC), Cambodia and Myanmar have cooperation with Rice Gene Discovery (Bio-Technology) Khampengsean, Kasetsart University, Thailand, Rockifeller and GCP
Using Bio-Technology in rice breeding program to capture quality into higher-yielding background

7. Objective To improve TDK 1 with aroma trait by using
Marker Assist Selection (MAS)

8. Material and methodology
Plant material:
The marker assisted backcross breeding is conducted using:
Recipient: TDK 1 (improved var. high yielding,..)
Donor: Homnangnouane ( Traditional var., aroma …)
HNN
TDK 1

9. How to transfer the aroma to
TDK 1?
Hom Nang Nuane (HNN) a Laos traditional variety, which carries the gene responsible for aroma to TDK 1 and then backcrossed to the recurrent parent generation until BC3F1 then selfed to BC3F3

10. Scheme of rice breeding
TDK1 x Homnangnouane
F2 x TDKI
BC1F1 x TDK1
BC2F1
BC2F2
MAS
BC2F3
RF, BIOTEC, KU
NAFRI
GCP, BIOTEC
BC3F1
BC3F2
Field Testing and trait validation of BC3F3
in NAFRI
Nov. 2007
May. 2008
Oct
May. 2007

11. Methodology DNA extraction DNA concentration PCR amplification
Lines selection

12. Polymerase Chain Reaction Polyacrylamide Gel Electrophoresis
DNA marker and PCR
The leaf samples of each BC generation were collected from ARC
DNA of both parents and backcrossed lines were extracted using the methods described:
At Khampengsean
Campus
Dellapota et al. (1983)
Fulton et al. (1995)
Rice Tissue
DNA extraction
Polymerase Chain Reaction
Polyacrylamide Gel Electrophoresis
Data Scoring

13. Parental Polymorphism Survey
Markers
Traits
Chromosome
TDK1
HNN
KDML105
BADH
Aroma 8
400
392
Waxy
Amylose content 6
413
RM204
212
RM21
Gel.tem. 11
252
249
RM457
417 No
RM5349
125
113
231
RM6897
141
151
RM190
251
RM597
336
RM8114
PSS-II-2
370
369
RM121
465
463
GT11
280
282
SNP3
Gel.tem. (Specific)
Mono

14. SNP3 for parental screening (Agarose gel)
TDK1
Homnangnouane
KDML105
Standard

15. Polyacrylamide gel of parents with markers: BADH, RM21 and RM5349

16. Markers used for progenies selectionNo
Markers
Traits
Chrom.
TDK1
HNN 1
BADH
Aroma 8
400
392 2
RM21
gelatinization temperature 11
252
249 3
RM5349
125
113

17. Number of plants and selected plants

18. Maturity, yield, plant height and pan./hill of 13 lines.No
Pedigree
Maturity
yield
Plant height
Pan./hill
(days)
(kg/ha)
(cm)
 (pan.) 1
TDK
135
3814 98 9 2
TDK
140
3385 3
TDK
3238 96 8 4
2977 5
TDK
3586 6
TDK
3336 7
TDK
3112
TDK
3220
142
TDK
3160 93 10
TDK
3113 11
TDK
3444 12
TDK1
3192 97 13
HNN
(Oct.15)
2689
138
5% (LSD)
0.23
359
5.82
1.05
CV(%)
11.6
4.8
10.90

19. Quality test

20. Maturity, yield, plant height and pan./hill of 3 lines.

21. TDK303-140-3-93-4-27 (V3) compare to TDK 1
Advantage:
RD6 aroma 32 38 39 45 46 3 5
- Aroma (TDK 1 No aroma)
Yield is semilar to TDK 1:
3-5 t/ha
Maturity is seminar to TDK 1: days,
Can grow in wet and dry season

22. TDK303-140-3-93-4-27 (V3) compare to TDK 1
Advantage:
Moderate resistance to BL, BPH,
Good adaptability to central southern parts
Respond to N
Good milling and eating quality
Disadvantage: not resistance to stem borer and gall midge

23. Grain characteristic of TDK303-140-3-93-4-27 (V3)

24. Grain shape and milled quality

25. Conclusion
Successful in first time using MAS backcrossing in rice breeding in Lao PDR
So TDK 1 aroma (TDK (V3) is a first improved aroma variety by using
MAS is good and accurate tool for breeding, especially for the recessive trait
Give more knowledge and experience for Lao staffs

28. Other alleles of BADH2? N N Y
Shi et al – deletion in exon 2.

29. Acknowledgement
NAFRI
Bio-tech Khampengsean KU
Lockifeller
GCP
ARC

30. Thank you