634 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /-, No.+,, (,**0) 38 PCR Bienvenido O. Juliano* * Philippine Rice Research Institute Cultivar Id

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1 634 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /-, No.+,, (,**0) 38 PC Bienvenido O. Juliano* * Philippine ice esearch Institute Cultivar Identification and Palatability Estimation of +. Typical Philippine ice Cultivars by the PC Method Sumiko Nakamura, Keitaro Suzuki, Kazutomo Haraguchi, Yoko Takemoto, Bienvenido O. Juliano* and Ken ichi Ohtsubo National ood esearch Institute,,++,, Kannondai, Tsukuba, Ibaraki, -*/20., *Philippine ice esearch Institute, Maligaya, Munoz, Nueva Ecija, Philippines We have developed five PC-based DNA markers for rice blast resistance. All the DNA markers developed in the present study were shown to be located on the same chromosomes as the target blast resistance genes and are estimated to be closely linked with them. Using PC for these primers, four typical Philippine rice cultivars were found to have di#erent DNA patterns. urthermore, we have developed a novel primer closely related to soluble starch synthase (SSS). This primer revealed marked di#erences between Japonica and Indica subspecies of rice. In addition, the results of PC were highly correlated with the amylose content. This is due to the close location of granule-bound starch synthase (GBSS) and SS on rice chromosome six at an approximate map distance of /cm. The PC results for +. typical Philippine rice cultivars also showed high correlation for the amylose content. The present authors developed a formula for estimating the amylose content and gel consistency by multiple regression analyses based on the results of PC as the explanatory variables. (eceived Apr.,+,,**0 ; Accepted Sep.,2,,**0) 0 + PC,0 DNA DNA 1 DNA 2 -*/20.,,++, * -++3 Maligaya, Munoz, Nueva Ecija, Philippines Corresponding author kenohtsu@affrc.go.jp 3 +* ++ PC DNA +1 Pia, Pii, Pita-,, Piz, Pik-m STS +, +- Goto +. Miyamoto +/ +0 Y. ukuta +1 QTL +,,**, STS 02 +.

2 39 : ao+,.o soluble starch synthase ; SSS ao+,0o branching enzyme ; BE ao+,0o debranching enzyme ; DBE +3,* STS PC 12 PC,+,, DNA +,**, +. No. + No. +- No. + PSBc+.,No., PSBc+2,No.- PSBc,2, No.. PSBc/., No. / PSBc0., No.0 PSBc00, No.1 PSBc1., No.2 PSBc12, No.3 PSBc2*, No.+* PSBc2, No.++ I0*, No. +, I0., No.+- I1. No. +. NSICc+*. ++ KDL +*/, + 3* IM-+**, CTAB DNA 02 3* *.. g CTAB DNA DNA - STS d APD +, A0/ 02,. APD DNA STS e Piz,1 SNP APD PC STS f +, J0 +. APD STS l SSS,3-* SSS 02+, DNA 02 EASY TAP TM Ver., DNA Invitrogen TOPO XL Cloning Kit 02 QIAGEN QIAprep Spin Miniprep Kit DNA ABI Big Dye Termination. Cycle Sequencing Kit PC Applied Biosystems ABI PISM -+* PC ABI Gene Amp PC System 31** TaKaa PC Thermal Cycler Dice PC +,. DNA DNA DNA DDBJ DNA Date Bank of Japan IGSP International ice Genom Sequencing Project http : //rgp.dna.a#rc.go.jp/cgi-bin /statusdb/irgsp-status.cgi Web : Gramene http : //www. gramene.org/ db/cmap/map-details / STS PC PC / +

3 636 /- +,,**0 +, 40 + STS a b c d e f g h i j k l m n o CCGCAGTTAGATGCACCATTAGAATTGCTTCATTGCCTGTGGA CCGCAGTTAGATCAAGTGGCAAGGTTCCATGTTTGGACTCAA TACCTGAACCAGCAAGCATGCGCG TACCTGAACCAGTATAATCTTTG CTCTCGTTTGTCTTTGTGTGTGCTG ACCAGCATTCACCCCCGAGCCC AGCGCGGCAAAATCAAACGTAATTGGC AGCGCGGCAAAACACGAACCGATGTTTC GCATTTTTAGCTATGAATCTGGAT TGTAGAATGAGGTGAGTTATTAACA GTCGGAGTGGTCAGACCGGCACAACTA GTCGGAGTGGTCAGACCGACAACTAG ATATTTGTGGAGAACCTCATCCCC TAGCTTTTGGTGATGGGCTCAGGC TGGCCGGCATGACTCAC ACTGGCCGGCATCAAGAC ACAACGCCTCCGATGAT ACAACGCCTCCGACAACAAG CCAGCTGTACGCCTGTACTAC CCAGCTGTACGTCTTCCCCAGC AAGCATGATCGCGTTATCCCAGGTG AAGAGCAGAAAGTAGAGATCACG GCGCCCGCTGGCACCGGC CAACCATCACACGATGACCACGAAGAG GAGACCGATATGCGATTC GTGGTGTTTAGATCCAGAGACTTAAC GTCGGAGTGTCAGACCGCGTGCCGTCCTCG GTTCGGCATTGGTCAGACCGGCAGGCAG TTTATCGATGGAGTTGATTTTGT TGTATCACAAGGACAGAGCGAG, +. PC / pmolel DNA.** ngl a b c d e f g h i j k l m n o :*40 l :*4/ l :*4. l :*4- l :*4/ l :*42 l :*4. l :*4+ l :*4. l :*4, l :*4/ l :*42 l :*4- l :*4- l :*40 l : *40 l : *4/ l : *4. l : *4- l : *4/ l : *42 l : *4. l : *4+ l : *4. l : *4, l : *4/ l : *42 l : *4- l : *4- l : *40 l , 0, 00 1* , 0, * 02 + l, l + l + l + l, l + l + l + l + l + l, l + l + l + l ABI Gene Amp PC System 31** *DNA,0* nm

4 41 : Juliano,+ Potato amylose 0 2 CAGAMPANG,,,- +** mg +/* mm *.*,/ wv 3/ *., ml *.,,/ moll, ml 0* -* +/ +,* 3 PC PC + * Microsoft Excel,**, Juliano,+ 0.* * cm PSB c+. -,40 +,42 PSB c+2, / PSB c,2,/4- +.4/ PSB c/.,24/ +.4- PSB c ,4/ PSB c00-04, +-4/ PSB c ,4/ PSB c PSB c2*,/40 +/4+ PSB c2,,041 +/4/ I0* *4/ I0.,340 +, I / NSIC c+*. +34* -, +-,,,- *., moll *.,,/ moll +- - r* DNA + DNA a, b, c +, d, e + Pii Pii 3,0.1* cm -...* cm,/ a Accession number AP**/2++ PAC P*1*0E * * cm --.** cm a Pii +, + A Pita-, Pita-, +,

5 638 /- +,,**0 +, 42 + A : 3,0.** cm-/.** cm Pii a B : +, /*.** cm0-.* cm Pita-, b, c C : * cm++0.* cm Pik-m d D : 0 /0.* cm1+.* cm Piz e //..* cm /3.** cm,0 b Accession number AL11,.+3 BAC OJ +++/-G*, 33./ +, /*..* cm c Accession number AL2./-.- BAC OSJNBa**,2**3 22./ +, 0,.,* cm b c Pita-, +, + B Pik-m Pik-m LP +/* , cm d Accession number AC +-.3,, BAC OSJNBa***.D* ,.3 cm++... cm d Pikm + C Piz Piz 0 0-.** cm02.0* cm,1 e Accession number AP**/0/3 PAC PO0.3 C++ 33.** 0 /0.-* cm /2.1* cm e Piz + D, f Accession number AP**/.., BAC OSJNBa**1-+*/ 3/.** 2.,.3 cm...0* cm g Accession number AP**.-+1 PAC PO00-E+* 3,.** + +*-.1 cm+*0., cm i Accession number AP**/1-* BAC OSJNBa**1-J+3 32.** cm J Accession number AL3,211. BAC OSJNBa**12*+1 32.** +, 00.0 cm01., cm. +. PC +, PC PC,. a e

43 : 639. +. PC a b c d e f g h i j j j k l m n o + 0+ 20* 21* 21*,** 12* 101 2,1 +,2* 2*/ 1.* 01* -13 + -1* 2+2 0** 2*2 +4 PSB c+.,4 PSB c+2-4 PSB c,2.4 PSB c/. /4 PSB c0. 04 PSB c00 14 PSB c1.

6 43 : PC a b c d e f g h i j j j k l m n o * 21* 21*,** 12* 101 2,1 +,2* 2*/ 1.* 01* * 2+2 0** 2*2 +4 PSB c+.,4 PSB c+2-4 PSB c,2.4 PSB c/. /4 PSB c0. 04 PSB c00 14 PSB c1. 24 PSB c12 34 PSB c2* +*4 PSB c2, ++4 I0* +,4 I I NSIC c+*., +. PC aj ++. M:DNA Marker. + : PSB c+., : PSB c+2 - : PSB c,2. :PSB c/. / : PSB c0. 0 : PSB c00 1 : PSB c1. 2 : PSB c12 3 : PSB c2* +* : PSB c2, ++ :I 0* +, :I0. +- :I1. +. : NSIC c+*.

7 640 /- +,,**0 +, 44 - SSS a:/ b:- G exon+ PC +. PSB C+., PSB C,2, PSB C0., I0.. f PC +. PSB C00, I1. 0 g STS g PC L- + 2 QTL +,2 g + +*-.1 cm+*0., cm +. PC / PC +. PSB C+2, PSB C/., PSB C1., PSB C2*, I0*, NSIC C+*. 0 +, j STS PC - j PC PSB C12, PSB C2,, +. h STS i STS DNA PC, a h i j i PC k, l, m, n, o ao+,0o debranching enzyme ; DBE pullulanase k 2 PC,3 2 DBE pullulanase SSSa,* k +. PC. I0* I0. PSB C DBE SSS,3-* SSS - a /, b - 2 NSS+ SSS -* NSS

45 : 641. +/ SSS l PC ++. M:DNA Marker. + :, : - :. : / : 0 : 1 : 2 : 3 : +* :KDL+*/ ++ : +, : +- : +. : +/ : PC 2 L- +.++ G exon + SSS precursor exon + -* l PC. No. + No., No. - No.. No. / No. 0 No.

8 45 : / SSS l PC ++. M:DNA Marker. + :, : - :. : / : 0 : 1 : 2 : 3 : +* :KDL+*/ ++ : +, : +- : +. : +/ : PC 2 L G exon + SSS precursor exon + -* l PC. No. + No., No. - No.. No. / No. 0 No. 1 No. 2 No. 3 No. +* KDL+*/ No. ++ No. +, No. +- No. +. No. +/ No3 No No., +1.. No No No. / +2 No No. 1 -, No No. 3,+.. No. +* +3 No. ++,* No. +,,3./ No. +-,. No No. +/ -,.+ PC + l +. PC. PC / GBSS granule bound starch synthase GBSS SSS 0 /cm -* SSS 2 branching enzyme m n +. PC. 2 SSS / - - / o +. PC.. k DBE l SSS m BE ng o SSS PC - r*.1/ r *.2. + / A B Pii, Pita-,, Pik-m, Piz / PC , +- DBE SSS BE SSS PC + r*.1/ r*.2. -,

9 642 /- +,,**0 +, 46 / k l m n g o PC A : PC y*./.-/3.x/.32/3.+ B : PC y*.033*.+x /+/. +332, APD * APD.0,0,, DNA.2 -*0-+*,**+ / 12 +* ,**. 0 PC ,**, 1 DNA /* +,,+-,,**- 2 PC ,**. 3 D /, +* No *1+ +32* ++ 1,3+ +32/ +, BL DNA ,**0 +- Pb+ 1 1/2/,**/ +. Goto, I., Jaw, Y.L., Baluch, A.A., Genetic studies on resistance of rice plant to blast fungus,iv, Link-age analysis of four genes,pi-a, Pi-k,Piz and Pi-i, Ann Phytopath Soc Japan,.1 (,),/,,/ / Miyamoto, M., Ando, I., ybka, K., Kodama, O., Kawasaki, S., High resolution mapping of the Indi ca-derived rice blast resistance genes.i. Pib, Mol, Plant Microbe Interact, +, QTL ukuta, Y., Yanoria, M. J.T., Ebron, L.A., Mercado, D.M. and Khush G.S., Genetic and breeding analysis using molecular marker,,,.eaction pattern of QTL for Philippines blast Isolate, Breeding esearch, - (Suppl),.0,** ,*,/ Baba, T., Nishihara, M., Mizuno, K., Kawasaki, T., Shimada, H., Kobayashi, E., Ohnish, S., Tanaka, K. and Arai, Y., Identification cdna cloning and gene expression of soluble starch synthase in rice (Oryza sativa. L)

10 47 : 643 immature seeds, Plant physiol +*-, /0// ,* Nakamura, Y., Towards a better understanding of metabolic system for amylopectin biosynthesis in plant : rice endosperm as a model tissue, Plant Cell Physiol,.-, 1+21,/,**,,+ Juliano, B.O., A simplifired assay for milled-rice amylose, Cereal Science Today, +0, +*, --.-.* +31+,, Cagampang, G.B., Perez, C.M. and Juliano, B.O., J. Sci. ood. Agric.,,., +/23+/ , /*2/+- +33,,. Williams, J.G.K., Kubelik, A.., Livak, K.J. and Tingey, S.V., DNA polymorphisms amplified by arbitrary primers are useful as genetic markers, Nucleic Acids esearch, +2, 0/-+0/-/ +33*,/ Pii / + +*2,**-,0 Jia. Y., Wang, Z., Singh, P., Development of dominant rice blast Pi-ta resistance gene markers, Crop Science,.,,,+./,+,**,,1 Hayashi, K., Hashimoto, N., Daigen, M., Ashikawa I., Development of PCbased SNP markers for rice blast resistance genes at the Piz locus, Theor Appl Genet, +*2, +,+,+,,*,**.,2 DH QTL.2, +33,,3 Jiang, H.W., Dian, W.N and Wu, P., Oryza sativa (indica cultivar-group)putative soluble starch Synthase I gene, NCBI ACCESSION AY,33.*.,**- -* Tanaka, K., Ohnishi, S., Kishimoto, N., Kawasaki, T. and Baba, T., Structure organization and chromosomal location of the gene encoding a form of rice soluble starch synthase, Plant Physiol, +*2, / -+, pp. +.2+/ , - / +1,**- +2.,+ +2 3,2

Differentiation and search for palatability-factors of world-wide rice grains by PCR method Sumiko NAKAMURA, Hiroshi OKADOME, Koh-ichi YOZA, Kazutomo

Differentiation and search for palatability-factors of world-wide rice grains by PCR method Sumiko NAKAMURA, Hiroshi OKADOME, Koh-ichi YOZA, Kazutomo HARAGUCHI, Tomoya OKUNISHI, Keitaro SUZUKI, Hikaru