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유전� 좌위 분�마커를 �용한 아밀로스 함량별 벼 품종 분류

�소명, 권�호, 강주�, �지윤, 조수민, 신�진, 차진경, 박�수, 조준현, �종�*

Classification of Korean Rice Cultivars through Amylose Content Using Molecular Markers Targeting the Waxy Locus

Korean Journal of Breeding Science 2021;53(4):361-372.
Published online: December 1, 2021

�촌진�청 국립�량과학� 남부작물부 논�용작물과

Paddy Crop Research Division, Department of Southern Area Crop Science, National Institute of Crop Science, RDA, Miryang, 50424, Republic of KoreaÿØÿàJFIFddÿìDuckydÿáqhttp://ns.adobe.com/xap/1.0/ ÿîAdobedÀÿÛ„ÿÀŠÿÄ     ! 1AQa"q� 2#w8ð‘¡ÁÑB3´µ6v¶7×X±Rr²$˜9áñb’C³Ãt%u&–ÖW·‚s'(xy¢ÂÓ4T”5f‡—ÇH¨¸Ø  !1AQaq"�‘¡2B ±ÁRbr#³´u67ðѲ3sTt5v8áñâSc$4Ä‚’¢C“ÔÔ%UÕÂÒÓ…•FVÅÿÚ ?ßàêÓ_A÷ú‹× ­j- ŠHÌûû>‚>Ì,šm*>½ôßÈÞfzp"áõö—£Téõ‹£—Ær÷Kkr¾ÒÙî��âœ^× »Òr.|_&€ã]¿|*œ™v¿P¶²uܶv”o‡±¿QÇ1Ÿ ¾�ŒŸmÍëw¾VJUöhu-ãIð"=L‹‡n§õ‘iù½AÆ•8Úö�"ûÛ²½þ„� ák�*Ò¿[yùu:ý.Ž‚v«U¤ÒÚQ+•)Ñá%F� D•Hy²V‡è×B훩kš>‘Hêy8øÓj©Ý¹ q~§9Dúñ4íKR‚þm±zõÚíQ„¥ßö)µ^Ê”Ó.J%k_tVi5NTê¥Ójg!'¡k©Çy|5as…¨ðÃOÏÁÈ»)R–¯Û¸ß©FMÔ¿3LÕ4çj¹6dÖùÚœ#êNIwUF©•]J ûq¡BŒô¹/(ËF™aãŠâfJöRzq3à\GÕ›™ ?ö~\£ 6å)6èý•Wµµ4¶m[O›ö^L0ìEË&r¤RMïØ¶*º×ÑC .ÍÂ]Un·l¥Š-1— 1ßr#Rj/¥´ø¯&QɈ׉˩·ás6­Rj=5Tgæ.´êy™.·PÓ¡:ÖJJíS«…÷:îßC8èÝ-Äù„2u]–d�&¢é»Òvøª·Uºz;¾»7p·9‘˜» 5Všã‰nLÖ¢µ£”¢"y)Ц">Ói×™Ñ(ûIDDd|œ¹Ö Yj0·­; L›§ãR¯fÙÞàÝS�˜:k¥ÚÃñtYäK%*ÒN2^ômñ|&dÄŸthó":‡ãÊe§Øy¾)u§P—¶ôQó¡ZöðW)gcC3Pv&“M¾ÇéM¯„ÂWìÏì¬d&¯Å´ãÛ²½�mvŸT¹èRoÞÕªM0Ü3ä*…F3Yº›dûèæãýÃ6¡®èÚ\8õ,¼\h÷Ý» kß”‘õâi¶¡<k× žÞNTõð¦wSëÔªº­Ñmljö•’j[>->±p£tU©Ôâ�%¯'§L‹R>&EÇBïôH$ßÝý�§ÉMQAU¶x[›$Z6’ÒÍv®Ôi&’_“a.K˜IQ{äh©yÆ’½ ’j"JùOCá ê×9òö—ìeäFŒªÒ�´ÞÿfŽjµîí;™ËšÎ©<[3”_m%ÃïÑ÷ ·•l‹¦Q@¥Ö4œ³Ñg²ü�=5$°™(J]YëÁ®c-OMqé<÷ÊÚõÇ�‡“ wSø²�¨¹z’œ«ïÚ—)kš$ÜË7ªV©I¥ïÅPý붾ͯnV+卵*t]i‚ÐŽD¥31ñí~S¨Añí1ûéÃCÐ2u)ÑÂʼnΛãQn-UÅoîi÷3ò´�:®«gß»rI8ý¯ÊØ“Wm*G zà«Ü•)ZÌ×§J}Åê§Yµ Y“lGeJ6ØcB2I ‹NS3×Qµ>kÔõÞ=KT›½ÅBëœâ—ÆÙÎT]ŠŽž‚W鿇¢ÂÞ6œ¬«Ü+ãSÓÛ¦ööï÷Ož—X©QgG©R¦? 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*Corresponding Author (E-mail: ccriljh@korea.kr, Tel: +82-55-350-1168, Fax: +82-55-352-3059)
• Received: August 1, 2021   • Revised: November 1, 2021   • Accepted: November 1, 2021

Copyright © 2021 by the Korean Society of Breeding Science

This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  • A total of 285 Korean rice cultivars were used for amylose content and genetic analyses using KASP markers related to the amylose content. We used three polymorphic KASP markers targeting the Waxy (Wx) gene and compared the genotyping efficiency with previously reported molecular markers. The rice cultivars were raised in Miryang, Korea, for one period. Genetic analysis of the rice cultivars revealed that the three KASP markers on the Wx gene were functional for genomic selection of SNPs related to amylose content variation. Therefore, the three KASP markers targeting different Wx alleles can be used for genomic selection. In addition, KASP markers were more efficient than previously reported molecular markers that target the same alleles. These findings can be beneficial for the breeding and genomic selection of high-quality rice in Korea.
쌀� 주요 구성성분� 단백질, 지질, 수분, 그리고 전분�다. 전분� 백미� 경우 중량 대비 76.7~78.4%를 차지하고 있다. 쌀� �미 � 조리특성� 단백질 함량, 아밀로펙틴 분지사슬 길� 분�, 아밀로스 함량, 호화특성 등� 따� 달�지며, � 중 가장 � �향� 미치는 것� 아밀로스 함량�다(Bao et al. 2006, Cagampang et al. 1973, Chen et al. 2006, Fuller et al. 2009, Hansen et al. 2002, Huang et al. 1998, Juliano 1985, Lee et al. 2016, Lestari et al. 2009, Little 1958, Ramesh et al. 2000).
쌀� 아밀로스 함량� 벼 6번 염색체�� Waxy (Wx) 유전�� 유전�형� �해서 찰벼, 저아밀로스, 메벼 � 고아밀로스로 분류�며, 아밀로스 합성� Granule Bound Starch Synthase 1 (GBSS1)� �해 조절�다(Sano 1984, Wang et al. 1992, Wang et al. 1990). Wx 유전�는 벼 재배종� 따� 여러 하플로타입� 존재하는�, �반 메벼� 하플로타입� Wxa, Wxb, Wxin� 야�벼형 대립유전� Wxiv�서 파��었다(Zhang et al. 2019). 고아밀로스 품종� Wxa 대립유전�를 가지고 있으며, Wx 유전�� 발현� Wxb 대립유전�를 가진 품종보다 10배 정� 높고, 아밀로스 함량� 25% ��� 차지한다. 아밀로스 함량� 16-23% 전후� �반 메벼 품종 중 ��니카 품종� Wxb 대립유전�를, �디카 품종� Wxin 대립유전�를 가지고 있다(Kumar et al. 1987). 찰벼는 Wx 유전� 2번 엑�� 23 bp� 중복 염기서열� �해 전사과정� �� 종결�는 wx 대립유전�를 갖고 있어 아밀로스 함량� 낮다(Isshiki et al. 2001, Teng et al. 2012, Wanchana et al. 2003). 한편, 찰벼와 멥쌀� 중간 정� 아밀로스 함량� 저아밀로스 품종 밀키퀸� 고시히카리� MNU (N-methyl-N-nitrosourea)를 처리해 만들어졌다(Ise et al. 2001). 밀키퀸� Wx 유전�� 4, 5번 엑�� SNP �연변�가 �겨 아밀로스 함량� 낮아졌으며 � 대립유전�는 Wx-mq로 명명�었다(Ise et al. 2001, Sato et al. 2002, Suto 1996).
한편, 분�마커를 �용하여 벼� 아밀로스 함량� 구분하기 위해 많� 연구들� 진행�었다. Bligh et al. (1995)� Simple sequence repeat (SSR) 마커� RM190를 �용하여 (CT)n 반복수� 따� 아밀로스 함량� 구분� 가능하다고 하였다. Yamanaka et al. (2004)� Wxa 대립유전�를 �정할 수 있는 Derived cleaved amplified polymorphic sequences (dCAPS) 마커를 보고했다. � 마커는 활용 시 EcoT14I 제한효소를 �용하며 제한�리 유무� 따� Wx 유전�� 첫번째 �트론 5’-�어맞추기부위(splice junction)� 위치한 G/T Single nucleotide polymorphism (SNP)� 검출하여 Wxa와 Wxb 대립유전�를 구별할 수 있다. �한 Yamanaka et al. (2004)� 찰벼 대립유전�� wx� 23 bp 중복염기서열� 대�으로 InDel 마커를 개발하였으며, Sato et al. (2002)� 밀키퀸� Wx-mq 대립유전�가 가진 SNP 선발� 위한 tetra-primer를 보고하였다. Shao et al. (2020)� 여러 Wx 대립유전�를 구분 가능한 6개� Kompetitive allele-specific PCR (KASP) 마커를 보고했다.
본 연구�서는 분�육종 기반 고품질 � 가공용 벼 품종� 다양화를 위해 Wx 유전� �� KASP 마커를 �용하여 국내 벼 품종 285개� 대한 유전�형� 파악하였다. �한 해당 마커를 �용하여 Wx 대립유전�형별 찰벼, 저아밀로스벼, 고아밀로스벼 품종 구분� 가능한지 검토하였다.
시험재료 � 아밀로스 함량분�
시험재료는 국내 육성 벼 품종� 오대벼 등 ��니카 품종� 253개, 태백벼 등 통�형 품종� 32개를 사용하였다. 2020년 국립�량과학� 남부작물부(경�남� 밀양)�서 보통기 보비재배법� 준하여 재배하였으며, 5월 6� 파종하여 29�간 육묘한 뒤 �앙� 6월 4�� 재�거리는 30×15 cm 간격으로 주당 1본씩 한 품종당 4열씩 심었다. 아밀로스 함량 분�� 위한 시료는 출수�태형� 따� 조�종, 중�종, � 중만�종� �� 출수 후 45�, 50� � 55�� 수확하였다. 아밀로스 함량� Juliano� 비색검정법(Juliano 1971)� 따� 분�시료를 전처리 한 후 UV-VIS spectrophotometer (UV-2700, Shimadzu Co, Japan)를 �용하여 측정하였다. 쌀가루 0.1 g� 1 mL �탄올과 9 mL 1 N NaOH를 가한 후 10분간 항온조건�서 호화시킨 샘플� �류수를 �해 용량� 100 mL로 맞췄다. 그 중 5 mL� 취하여 1 mL� acetic acid, 2 mL� 2% I-KI (iodine solution)� 가한 샘플� �류수를 �해 용량� 100 mL로 맞춘 다� 20분 후 620 nm 파장� 대한 �광�를 측정했다. � 값� 표준검량�� 대입해 아밀로스 함량� 구했다.
Genomic DNA 추출
Genomic DNA는 벼� 어린 잎� 채취하여, CTAB 방법(Murray & Thompson 1980)� �용하여 다�과 같� 추출했다. 벼 어린 잎 3~4 cm와 3 mm 마쇄용 구슬(Masuda, Cat, 12-410-032) 1개를 2 mL Eppendorf tube (Axygen, MCT-200)� 넣고 액체질소� 5~10분간 침�시킨 후 1분간 vortex했다. 마쇄� 잎 샘플� 700 µL� DNA Extraction buffer (Biosesang)를 넣고, 65℃로 30분 처리했다. 마쇄� 잎 샘플과 DNA Extraction buffer 혼합물� 700 µL CI 용액(chloroform:isoamylalcohol=24:1)� 첨가하여 10분간 �반한 후 13,000 rpm으로 15분간 �심분리하여 500 µL �등액� 1.5 mL Eppendorf tube� 옮겼다. �등액� 1000 µL� 90% ethanol� 첨가하여 2~3회 혼합한 후 -20℃로 2시간 처리한 뒤 13,000 rpm�서 10분간 �심분리하여 �등액� 버리고 침전물� 70% ethanol로 세정 후 건조했다. 추출� DNA는 TE buffer (10 mM Tris-HCL, pH 8.0; 2.5 mM EDTA)� 녹여 PCR� �용했다.
유전�형 분�
유전�형 분�� 위해 KASP마커 � InDel, CAPS 마커를 사용했다. KASP 마커는 모부본 다형성� 분�하여 다형성� 보� 아밀로스 함량 관련 마커 3개를 �용했다(Shao et al. 2020). PCR 플레�트 한 well당 KASP-TF V4.0 2×Master Mix (LGC Genomics, London, UK) 4.86 µL, KASP by Design Primer Mix 0.14 µL� 1 µL당 DNA �량� 5 ng� ��� ��한 DNA 샘플� 5 µL 분주한 뒤 Cheon et al. (2018)� 보고한 바� 따� Touchdown PCR 방�으로 PCR했다. PCR 단계는 다�과 같았다. 우선 15분간 94℃로 유지한 뒤, 94℃ 20초, 61℃ 1분간 유지하는 2단계로 �루어진 과정� 10번 반복했다. �후 94℃ 20초, 55℃ 1분간 유지하는 과정� 26번 반복하고, 37℃�서 1분간 유지한 뒤 형광 측정 전까지 샘플 온�를 4℃로 유지했다. PCR 산물� Real Time PCR (QuantStudio 3, appliedbiosystems)로 형광� 측정 했다. InDel � CAPS 마커는 기존� 보고� Wx 유전� 관련 마커를 �용했다(Sato et al. 2002, Wanchana et al. 2003, Yamanaka et al. 2004). � 마커별 PCR 단계는 다�과 같았다. Glu-23 마커는 프��머를 10 pmole/µL ��로 0.5 µL씩 첨가하고, DNA 5 µL (50 ng/µL), dNTP 0.8 µL (0.2 mM), 10×buffer 1.5 µL, Taq 0.1 µL (1.25 unit), dH2O 6.6 µL를 첨가한 뒤 PCR 반�시켰다. Wx-mq 마커는 네 가지� 프��머를 모� 20 pmole/µL ��로 1 µL씩 첨가하고, DNA 2 µL (50 ng/µL), dNTP 1 µL (0.2 mM), Taq 0.2 µL (2.5 unit), dH2O 12.8 µL를 첨가한 뒤 PCR 반�시켰다. PCR 단계는 94℃�서 5분 처리 후 94℃ 1분, 69℃ 1분, 72℃ 2분 유지하는 3단계로 �루어진 과정� 28번 반복했다. �후 72℃�서 5분 처리 후 4℃ 냉장보관했다. Wx_a,b 마커는 프��머를 10 pmole/µL ��로 0.5 µL씩 첨가하고, DNA는 50 ng/µL ��로 5 µL, 0.2 mM dNTP 0.8 µL, 10×buffer 1.5 µL, Taq 0.1 µL, dH2O 6.6 µL를 첨가한 뒤 PCR 반�시켰다. 먼저 94℃�서 2분 처리 후 94℃ 30초, 55℃ 30초, 72℃ 1분 유지하는 3단계로 �루어진 과정� 35번 반복했다. �후 72℃�서 5분 처리 후 반�� 마친 PCR 산물� 제한효소 처리 전까지 4℃ 냉장보관했다. PCR 산물 20 µL당 5 µL� 비율로 EcoT14I를 첨가하고 37℃�서 하룻밤 처리했다. PCR � 제한효소 처리를 마친 샘플� 아가로스 겔�서 전기�� 한 뒤 �외선 조명� 비추어 촬�한 사진� 바탕으로 유전�형� 분�했다.
국내 육성 벼 285품종 아밀로스 함량 분�
벼 285품종 보통기 재배 시 백미 아밀로스 함량 �균� 17.9%�며 표준편차는 3.81�었다(Fig. 1, Table 1). ��니카 251품종 중 찰벼는 19품종, 저아밀로스벼는 5품종, 메벼는 223품종, 고아밀로스벼는 4품종�었고, 통�형 34품종 중 찰벼는 3품종, 저아밀로스벼는 0품종, 메벼는 22품종, 고아밀로스벼는 9품종�었다(Table 2). Bao et al. (1990)� 벼 전분� 신���계(RVA) 측정 결과는 벼� 외관� 아밀로스 함량(AAC)와 강한 �관관계가 있으며, 마커 유전�형 ��터와 함께 분�한 결과 Wx 유전�가 신���계로 측정�는 벼 전분� �화학� 성질� 좌우하는 주요 유전��고 보고하였다. 따�서 벼 전분 관련 형질 육종 시 Wx 유전�� 대립유전�형� 분�마커를 활용하여 미리 파악 가능하다면 아밀로스 함량� 수확기 전� 예측할 수 있기 때문� 보다 효율�� 육종� 가능할 것�다.
KASP 마커� ��
최근�는 KASP와 같� SNP 기반� 마커를 �용하여, Wx 유전�� 하플로타입� �욱 다양하게 �혀낼 수 있게 �었다. KASP 마커를 활용하면 전기��� 걸리는 시간� 줄어들어 �욱 효율�으로 서로 다른 Wx 대립유전�를 가진 품종 � 계통 구별� 가능하다. 형광표지를 �용한 비 전기�� 방�� 분�마커 중�는 Taqman과 KASP마커가 가장 보편�� 것으로 알려져 있는�, KASP마커는 Taqman마커� 비해 제작비용� 저렴하다(Yang et al. 2020). �한 Semagn et al. (2014)� �하면 KASP 마커는 다른 SNP 마커들� 비해 �균 오류율� 0.7~1.6% 정�로 낮� 편�고 한 가지� 마커 세트로 다양한 집단� 대해 �용� 가능해 필요한 마커 수가 �어 효율��다. 기존� 아가로스겔 기반� 마커는 PCR 후 전기�� 과정� 추가�으로 필요하기 때문�, 최근�는 대량 검정 � 분� 시간 단축� 위해 KASP, Taqman probe를 �용한 비 전기�� 방�� 분�마커 개발� 관한 연구가 활발히 진행�고 있다.
찰벼 품종 분류
Shao et al. (2020)� Wx 유전� 특��� KASP 마커 6개를 보고하였으며, � 중 K2-wx 마커는 2번 엑�� G/23N SNP를 구별한다(Table 3, Fig. 2). � 마커는 기존� 보고� 아가로스겔 기반 마커� Glu-23과 타겟� ��하다(Table 4). K2-wx 마커 검정 결과 23 bp 삽입� 가진 품종� 아랑향찰, 백설찰, 백옥찰, 보람찰, 보�찰, 보��찰, 청백찰, �진찰, 해�찰, 한아름찰, 한강찰, 한강찰1호, 화선찰, �진주찰, 진설찰, 조��찰, 눈��찰, �주찰, 설향찰, 신선찰, 운백찰, 운�찰로 � 22 개였고(Fig. 3A), 아밀로스 함량� 최저 6.5%, 최고 9.0%�며 �균 7.9%로 모� 찰벼 품종�다(Table 5).
벼� 찰성과 메성� 6번 염색체 Wx 유전�� 2번 엑� 내 염기서열� 중복� �해서 구분�다고 보고�어 있으며, � �역� InDel� 대�으로 Glu-23 마커가 개발�었다(Wanchana et al. 2003). 해당 마커는 ��니카형 � 통�형 찰벼�서 ��한 유전�형� 나타내었다(Table 6, Fig. 4A). �와 같� 결과를 통해 기존� 아가로스 겔 기반 분�마커로� 찰벼를 선발하는 것� 가능하다고 �단�다. 그러나 아가로스 겔 기반 분�마커는 검정 과정�서 PCR 후 전기�� 과정� 수행해야 하므로, KASP 마커를 활용하는 것� 편리하다. K2-wx 마커 염기서열� ��성� 분�한 결과 기존� 알려진 찰벼 대립유전� wx� 있는 23 bp 중복염기서열 �역� 대�으로 제작� 마커임� 확�했다.
밀키퀸 후대 저아밀로스벼 품종 분류
Wx 유전�� �연변�로 �해 저아밀로스 특성� 나타나게 하는 Wx-mq 대립유전�를 구별하기 위해 제작� YR-wxmq 마커는 4번 엑�� A/G SNP를 구별한다(Table 3, Fig. 2). YR-wxmq 마커 검정 시 Sato et al. (2002)� 보고한 마커와 마찬가지로 국내 육성 벼 285품종 중 밀키퀸 후대를 구별 가능했다. Wx 유전��서 해당 마커 위치� A SNP가 있어 Wx-mq 대립유전�를 가진 것으로 나타난 품종� 미호, 월백 � 품종�었고(Fig. 3B), �들� 모� 밀키퀸 후대 품종�다. � 품종� 아밀로스 함량� �� 12.3%, 13.6%�고 �균� 12.9%�었다(Table 5). 다른 저아밀로스 품종 중 백진주와 백진주1호는 du1 유전�� �해 저아밀로스 형질� 나타내기 때문� YR-wxmq 마커는 월백, 미호와 달리 G SNP를 가지는 것으로 나타났다(Table 6). �한 건양2호� 저아밀로스 형질과 밀접히 연관� SSR 마커는 선발�었으나 �� 유전�좌는 아� 명확히 �혀지지 않아 추가�으로 유전연구가 필요하다. 한편 아가로스겔 기반 마커 Wx-mq는 염기 치환 부위� 따� Wx-mq 대립유전� 특��� 유전� 단편� ����� 고안� 마커�다(Table 4). � 마커를 �용해 국내 벼 품종 중 밀키퀸 후대 저아밀로스벼 품종 구분� 명확하게 가능하였다(Fig. 4B). 그러나 Wx-mq 마커는 KASP 마커� 비해 검정시 필요한 프��머 수가 많고 전기�� 과정� 추가�으로 필요하여 시간 � 비용 측면�서 비효율��다.
고아밀로스벼 품종 분류
Shao et al. (2020)� 보고한 고아밀로스 품종 특��� 마커 K1-Wxa/Wxb를 �용하여(Table 3) 국내 육성 285개 품종� 유전�형� 분�한 결과 고아밀로스 대립유전� Wxa 특��� G SNP를 가진 품종� 청우, �담쌀, 고아미, 미면, 미우, 목우, 녹우, 팔방미, 새고아미, 새미면, �우 11품종으로 모� 고아밀로스 품종�었다. Wxa 대립유전�를 가진 품종들� 아밀로스 함량� 최저 23.9%, 최고 39.1%, �균 27.2%�었다(Table 5, Fig. 2). Wxb를 가진 품종� 오대벼 등 274품종�었다. 한편, 고아밀로스 품종으로 알려진 고아미2호와 신길� T base 대립유전�형� 가지고 있었다(Table 6, Fig. 3C).
따�서 고아미 2호와 신길� Wx 유전�좌� 위치한 고아밀로스 대립유전� 특��� SNP를 가지고 있지 않고 Wx 외� 다른 유전�� 변�� �해 고아밀로스 형질� 나타내는 것으로 �단�다. Butardo et al. (2012)� 고아미 2호는 Starch Branching Enzyme (SBE) 유전�가 결실� �해 아밀로스 함량� �가하는 amylose extender (AE) 유전�를 가지고 있는 것으로 보고한 바 있다. �한 신길� 기존� 고아밀로스 품종보다 아밀로스 함량� 약 2% 정� 낮� 특성� 가지고 있는�, �러한 특성� 나타내는 �� 유전�는 기존� Wx 유전�와 다른 염색체� 위치하는 것으로 �단�다. 위� 연구결과들� 종합하여 보았� 때, 고아미 2호와 신길� 고아밀로스 형질� 대해 유전분�과 유전� 지� 작성 등� 연구가 추가�으로 필요할 것으로 �단�다.
아밀로스 함량� 따� 벼 품종� �별할 수 있는 분�마커들� 다양하게 보고�어 있다. SSR 마커� RM190� (CT)n 반복수가 10개 �하� 경우 고아밀로스 품종으로 분류 가능해 아가로스 겔 �미지만으로 쉽게 품종 �별� 가능하다. 한편, 국내 육성 고아밀로스 품종� 고아미와 새고아미는 SSR 마커를 �용한 유전�형 분��서 �반 메벼와 구분� 불가능했다. �는 �들 품종� 유전� 배경� 잡초성 벼(weedy rice)� 김천앵미와 ��앵미�서 유래�어 기존� 알려진 �디카 유래 Wx 대립유전�를 가져 고아밀로스 형질� 나타내는 품종들과 다른 유전� 배경� 가지기 때문� 것으로 �단�다.
한편, Yamanaka et al. (2004)� 보고한 Wx_a,b CAPS 마커는 KASP 마커� K1-Wxa/Wxb 마커와 ��하게 Wx 유전�� G/T SNP가 타겟�다(Table 4). Wx_a,b 마커� 고아밀로스 벼 품종 구분� 가능하였지만(Fig. 4C), PCR 후 전기�� 전� 제한효소 처리가 추가�으로 필요하기 때문�, ��한 SNP를 타겟으로 제작� KASP 마커를 활용하는 것� 시간 � 비용 측면�서 � 효율��다.
본 연구는 국내 육성 벼 285품종� �용하여 Waxy (Wx)유전� ��서 다형성� 보�는 SNP 마커 중 아밀로스 함량 유전� 관련 마커 3개로 유전�형� 분�했다. 아밀로스 함량� �균� 17.9%, 표준편차는 3.82�었다. 고아밀로스 대립유전� Wxa 특��� K1-Wxa/Wxb 마커는 6번 염색체 1.77 Mb� G/T SNP를 구별하며, G base 대립유전�형� 가진 품종� 고아미 등 고아밀로스 품종�었다. 해당 고아밀로스 품종들� 아밀로스 함량 범위는 23.9%~39.1%�고, �균� 27.2%�었다. K2-wx 마커는 6번 염색체 1.77 Mb� G/23N SNP를 구별하며, � 중 Wx 유전�� 23 bp 삽입� 있는 wx 대립유전�를 가진 품종� 모� 찰벼로 구분�었다. 찰벼 품종� 아밀로스 함량 범위는 6.5~9.0%�고 �균� 7.9%였다. �한 Wx 유전� �연변�로 �해 저아밀로스 특성� 나타나게 하는 Wx-mq 대립유전�를 갖고 있는 품종� 미호, 월백 � 품종�며 아밀로스 함량� �� 12.3%, 13.6%, �균� 12.9%�었다. Wx-mq 대립유전� 특��� YR-wxmq 마커는 A/G SNP를 구분하며 � 마커로 A SNP를 갖고 있어 Wx-mq 대립유전�로 �해 반찰성� 나타내는 월백과 미호를 다른 품종과 구분 가능했다. 따�서 Wx 유전�좌 �� 3개� KASP 마커 모� 특정 범위� 아밀로스 함량� 가진 품종� �별 가능한 마커� 것으로 �단�다. �와 같� 결과는 국내 벼 품종 육성 시 아밀로스 함량 관련 유전체 선발� 유용한 정보가 � 것�다.
본 논문� �촌진�청 연구사업(과제번호: PJ01357203 �남지역�서� 국내 육성 벼 품종별 작물학� 특성 �가)� 지�� �해 �루어진 것임.
Fig. 1
Amylose content distribution of 285 Korean rice cultivars
KJBS-53-361-f1.tif
Fig. 2
Location of SNPs of each KASP marker on Waxy gene
KJBS-53-361-f2.tif
Fig. 3
KASP SNP marker genotyping result graph
KJBS-53-361-f3.tif
Fig. 4
Gel-based marker genotyping result
KJBS-53-361-f4.tif
Table 1
Amylose Content and Heading Date of 285 Korean rice cultivars
Table 1
Cultivars Ecotype Heading Date* Amylose Content (%)
Odae Japonica 8/3 17.6
Unbong Japonica 8/2 16.5
Geumo Japonica 8/4 17.5
Jinmi Japonica 8/6 16.6
Obong Japonica 8/2 17.7
Namwon Japonica 7/31 18.9
Jinbu Japonica 8/3 12.8
Jinbuol Japonica 7/25 16.4
Sinunbong Japonica 8/4 17.6
Sangju Japonica 8/4 17.3
Dunnae Japonica 8/3 17.8
Joryeong Japonica 8/4 18.6
Unjang Japonica 8/2 16.9
Samcheon Japonica 8/4 17.2
Junghwa Japonica 8/4 16.6
Keumobyeo2 Japonica 8/15 16.6
Guru Japonica 8/5 17.4
Sangjuchal Japonica 8/7 8.3
Undoo Japonica 7/30 16.6
Manan Japonica 8/5 18.5
Jeogjinju Japonica 8/4 17.0
Taebong Japonica 8/2 16.4
Jungsan Japonica 8/7 18.1
Saesangju Japonica 8/5 17.0
Manchoo Japonica 8/4 17.5
Manho Japonica 8/3 18.0
Namil Japonica 8/7 18.1
Joan Japonica 8/2 16.6
Heugkwang Japonica 8/13 18.3
Goun Japonica 8/1 16.6
Unkwang Japonica 8/4 17.6
Josaengheugchal Japonica 8/6 8.4
Odae1ho Japonica 8/3 17.1
Sinunbong1 Japonica 8/3 18.0
Manna Japonica 8/2 18.1
Keumo3 Japonica 8/2 18.8
Hwangkeumbora Japonica 8/4 19.0
Sandeuljinmi Japonica 8/5 18.2
Junamjosaeng Japonica 8/5 18.6
Pyeongwon Japonica 8/4 16.6
Unmi Japonica 8/4 16.9
Handeul Japonica 8/9 17.4
Jogwang Japonica 8/5 18.1
BoSeog Japonica 8/1 19.4
Joami Japonica 8/5 17.6
Hanseol Japonica 7/31 18.2
Joun Japonica 7/31 18.5
Geumyoung Japonica 8/4 17.7
Manjong Japonica 8/13 19.2
Wolbaek Japonica 8/4 13.6
Jopyeong Japonica 8/1 18.3
Jeogjinjuchal Japonica 8/7 7.9
Cheongbaekchal Japonica 8/2 8.0
Seolemi Japonica 8/5 17.5
Joeunheukmi Japonica 8/4 16.9
Saeodae Japonica 8/3 19.8
Jinseolchal Japonica 7/27 8.3
Jungmo1024 Japonica 8/4 18.0
Sanhomi Japonica 8/4 18.0
Hwawang Japonica 8/6 17.6
Nunkeunheugchal Japonica 8/6 8.3
Asemi Japonica 8/5 19.2
Ondami Japonica 8/5 18.0
jungmo1032 Japonica 7/30 18.9
Jinok Japonica 8/1 17.8
BaegIlmi Japonica 7/26 17.2
Asemi1ho Japonica 8/8 18.7
Haedamssal Japonica 8/6 19.7
Unilchal Japonica 8/7 8.9
Jungmo1043 Japonica 8/5 19.1
Jinhan Japonica 8/3 17.9
Jopum Japonica 7/30 18.0
Joil Japonica 8/6 17.6
Danpyeong Japonica 8/5 19.0
Unbaekchal Japonica 8/4 8.4
Jinkwang Japonica 8/6 17.1
Seongsan Japonica 8/7 18.6
Jonong Japonica 8/9 19.2
Shinpyeong Japonica 8/4 17.4
CW92MR Japonica 8/7 17.2
Haedeul Japonica 8/10 17.4
IS592BB Japonica 8/5 16.7
Sinseonchalbyeo Japonica 8/11 7.8
Giho Japonica 8/15 17.7
Hwaseong Japonica 8/16 17.9
Yeongdeog Japonica 8/14 18.5
Palgong Japonica 8/12 18.0
Hwajin Japonica 8/15 19.3
Janngan Japonica 8/12 18.3
Seoan Japonica 8/13 17.2
Hwayeong Japonica 8/14 17.2
Hwaseonchal Japonica 8/12 8.1
Gancheok Japonica 8/13 19.0
Hwajung Japonica 8/12 19.8
Daeripbyeo 1 Japonica 8/15 20.1
Nongan Japonica 8/16 16.6
Juan Japonica 8/13 17.3
Keumobyeo 1 Japonica 8/11 17.9
Naepung Japonica 8/10 17.4
Daejin Japonica 8/17 18.6
Sura Japonica 8/15 17.6
Hwabong Japonica 8/16 18.1
Jinpum Japonica 8/13 19.2
Sobi Japonica 8/15 17.8
Seolhyangchal Japonica 8/11 8.3
Sampyeong Japonica 8/15 18.6
Hwaan Japonica 8/14 19.4
Manpung Japonica 8/16 19.4
Haepyeong Japonica 8/12 17.2
Saegyehwa Japonica 8/18 18.9
Manwol Japonica 8/16 18.9
Yeongan Japonica 8/17 19.1
Geuman Japonica 8/18 18.5
Daepyeong Japonica 8/18 18.2
Samdeog Japonica 8/15 18.3
Pungmi Japonica 8/12 16.9
Boseogchal Japonica 8/14 8.0
Haepyeongchal Japonica 8/14 7.9
Seoan1 Japonica 8/17 19.4
Geunnun Japonica 8/17 16.5
Cheonga Japonica 8/12 18.4
Gangbaek Japonica 8/16 18.7
Haechanmulgyeol Japonica 8/14 19.2
Donghaejinmi Japonica 8/17 19.4
Cheongdam Japonica 8/15 19.3
Hongjinju Japonica 8/16 17.9
Cheongan Japonica 8/15 19.2
Borami Japonica 8/18 20.4
Heugseol Japonica 8/16 17.4
Haeoreumi Japonica 8/16 17.8
Boseogheugchal Japonica 8/16 9.0
Migwang Japonica 8/19 19.1
Cheongnam Japonica 8/15 20.1
Suryeojinmi Japonica 8/15 18.8
Gangchan Japonica 8/19 17.5
Suan Japonica 8/17 20.0
Shinbaeg Japonica 8/20 18.6
Dongbo Japonica 8/18 18.3
Jungsaenggold Japonica 8/15 16.2
Seonhyangheukmi Japonica 8/14 17.5
Daebo Japonica 8/15 17.1
Geonyang2 Japonica 8/18 11.3
Dabo Japonica 8/17 18.0
Dodamssal Japonica 8/17 39.1
Bodrami Japonica 8/17 19.6
Sinbo Japonica 8/19 18.8
Jungmo1034 Japonica 8/15 19.8
Haepum Japonica 8/18 19.2
Seonpum Japonica 8/16 17.7
Youngbo Japonica 8/15 17.6
Samkwang1 Japonica 8/16 17.2
Cheongpum Japonica 8/15 17.7
Sangbo Japonica 8/14 19.1
Heugjinmi Japonica 8/17 17.9
Boramchal Japonica 8/15 7.5
Jeogjinju2 Japonica 8/18 18.8
Dongjin Japonica 8/19 19.4
Daecheong Japonica 8/17 19.0
Tamjin Japonica 8/21 18.2
Kyehwa Japonica 8/20 17.5
Ilpum Japonica 8/18 18.2
Manguem Japonica 8/17 18.7
Hwanam Japonica 8/16 18.2
Daean Japonica 8/17 18.7
Yangjo Japonica 8/15 19.3
Hwasin Japonica 8/20 20.7
Hyangnam Japonica 8/18 18.4
Ilmi Japonica 8/18 18.7
Dongan Japonica 8/17 20.1
Hwasam Japonica 8/20 18.3
Daesan Japonica 8/21 19.4
Nampyeong Japonica 8/19 20.5
Heugnam Japonica 8/17 19.6
Aranghangchal Japonica 8/18 7.6
Hoan Japonica 8/19 19.5
Dongjinchal Japonica 8/16 7.6
Mihyang Japonica 8/17 19.5
Sindongjin Japonica 8/18 20.1
Sujin Japonica 8/21 19.1
Hojin Japonica 8/15 20.2
Heughyang Japonica 8/22 21.4
Junam Japonica 8/20 19.3
Goami Japonica 8/19 27.1
Dongin1 Japonica 8/20 19.6
Baegjinju Japonica 8/21 11.2
Seolgaeng Japonica 8/18 19.7
Goami2 Japonica 8/20 32.0
Manmi Japonica 8/18 13.6
Samkwang Japonica 8/19 19.3
Hopyung Japonica 8/19 19.5
Seopyeong Japonica - 19.0
Hwarang Japonica - 18.3
Gopum Japonica 8/17 18.8
Cheongho Japonica 8/16 20.2
Hanmaeum Japonica 8/16 18.9
Pungmi1 Japonica 8/10 16.7
Onnuri Japonica 8/17 19.3
Hanam Japonica 8/18 18.3
Dongjin2 Japonica 8/17 18.7
Baegjinju1ho Japonica 8/21 11.3
Hwangkeumnuri Japonica 8/20 19.4
Dami Japonica 8/19 19.7
Malgeumi Japonica 8/19 19.2
Hopum Japonica 8/17 18.4
Saenuri Japonica 8/19 20.2
Chilbo Japonica 8/17 18.4
Hwanggeumnodeul Japonica 8/19 19.3
Baegseolchal Japonica 8/16 7.4
Cheongcheongjinmi Japonica 8/17 20.5
Jinbaek Japonica 8/20 20.2
Dacheong Japonica 8/23 18.8
Jinsumi Japonica 8/19 19.3
Jungmo1006 Japonica 8/18 19.2
Deuraechan Japonica 8/19 19.1
Haiami Japonica 8/18 18.5
Baekogchal Japonica 8/21 6.5
Danmi Japonica 8/20 13.5
Cheonghaejinmi Japonica 8/19 19.3
Honong Japonica 8/21 19.3
Seomyeong Japonica 8/18 19.6
Younghojinmi Japonica 8/21 18.9
Jinbo Japonica 8/16 18.4
Boramchan Japonica 8/19 19.3
Anmi Japonica 8/16 16.9
Mipum Japonica 8/22 19.0
Chinnong Japonica 8/20 18.1
Geongganghongmi Japonica 8/20 20.1
Sodami Japonica 8/20 18.8
Sukwang Japonica 8/17 18.7
Huimangchan Japonica 8/16 19.0
Geonyangmi Japonica 8/18 19.7
SaeIlmi Japonica 8/19 19.7
Saegoami Japonica 8/18 27.1
Cheongun Japonica 8/18 19.4
Hyeonpum Japonica 8/20 20.5
Chindeul Japonica 8/22 17.9
Heugsujeong Japonica 8/19 18.9
Misomi Japonica 8/19 18.8
Anbaek Japonica 8/18 19.5
Manbaek Japonica 8/22 17.5
Saeshin Japonica 8/22 19.9
Cheonghyangheukmi Japonica 8/23 20.3
Sinjinbaek Japonica 8/21 19.7
Youngjin Japonica 8/20 20.3
Saechilbo Japonica 8/18 18.1
Miho Japonica 8/23 12.3
Aromi Japonica 8/20 18.1
MY299BK Japonica 8/17 18.0
Yechan Japonica 8/19 19.0
Saeilpum Japonica 8/17 17.8
Keunpum Japonica 8/15 19.3
Taebaek Tongil 8/10 17.9
Cheongcheong Tongil 8/19 17.0
Jungweon Tongil 8/12 19.3
Yongmun Tongil 8/10 16.7
Yongju Tongil 8/11 17.3
Dasan Tongil 8/12 17.0
Namcheon Tongil 8/13 17.9
Hyangmi2 Tongil 8/10 21.3
Anda Tongil 8/17 19.4
Areum Tongil 8/12 19.1
Hanareum Tongil 8/18 19.2
Dasan1 Tongil 8/14 17.3
Keunseom Tongil 8/18 17.4
Hangangchal1 Tongil 8/19 7.5
Dasan2 Tongil 8/14 18.6
Hanareum2 Tongil 8/15 17.2
Palbangmi Tongil 8/16 26.2
Mimyeon Tongil 8/13 26.6
Saemimyeon Tongil 8/20 26.6
Hanareumchal Tongil 8/19 6.8
Shingil Tongil 8/19 23.5
Hangangchal Tongil 8/19 7.5
Nampung Tongil 8/17 19.6
Jangseong Tongil 8/14 18.5
Hyangmibyeo1 Tongil 8/16 18.0
Segyejinmi Tongil 8/10 18.0
Hanareum3 Tongil 8/15 17.3
Hanareum4 Tongil 8/16 17.7
Geumgang1 Tongil 8/20 18.9
Mogwoo Tongil 8/29 25.3
Nokwoo Tongil 9/2 23.9
Yeongwoo Tongil 8/19 26.7
Cheongwoo Tongil 8/26 25.4
Miwoo Tongil 8/21 25.6

*Transplanted on 4th June, 2020

Table 2
Distribution of Amylose content of 285 Korean rice cultivars
Table 2
Ecotype waxy Low amylose Intermediate amylose High amylose
Japonica 19 5 223 4
Tongil 3 0 22 9
Table 3
KASP marker information targeting Waxy gene
Table 3
Marker Target SNP Primer Sequence
Allele X (FAM) Allele Y (HEX) Common
K1-Wxa/Wxb G/T TCATCAGGAAGAACATCTGCAAGG TCATCAGGAAGAACATCTGCAAGT GCCCAACACCTTACAGAAATTAGCA
K2-wx G/23N TTCCAGGGCCTCAAGCCCC GTTCCAGGGCCTCAAGCCCA CGCTGGTCGTCACGCTGA
YR-wxmq A/G GATGAACACACGGTCGACTCCAT ATGAACACACGGTCGACTCCAC GAGAGGGTGAGGTTTTTCCATTGCTA
Table 4
Gel-based marker information targeting Waxy gene
Table 4
Marker Name Chr. Sequence Annealing Temp. (℃) Restriction Enzyme Traits related Reference
Forward Reverse
Wx_a,b 6 TGTTGTTCATCAGGAAGAACATCTCCAAG TTAATTTCCAGCCCAACACC 55 EcoT14I, 37℃, 1 h~over night high amylose Yamanaka et al. 2004. Theor Appl Genet 108:1200-1204
Glu-23 6 TGCAGAGATCTTCCACAGCA GCTGGTCGTCACGCTGAG 60 waxy Wanchana et al. 2003. Plant Sci 165: 1193-1199
Wx-mq 6 GGTGAGGTTTTTCCATTGCTACAAGCA GCCCTGGTAGGAGATGTTGTGGAT 69 low amylose Sato et al. 2002. Breeding Sci 52:131-135
CAAGTACGACGCAACCACGGTAAG GGTGGACTAGACGATCTGGGTTCAA
Table 5
Cultivars classified by Waxy SNP marker genotypes and their amylose content distribution
Table 5
Marker SNP Cultivar Amylose Content (%)
Min. Max. Average
K1-Wxa/Wxb G Cheongwoo, Dodamssal, Goami, Mimyeon, Miwoo, Mogwoo, Nokwoo, Palbangmi, Saegoami, Saemimyeon, Yeongwoo 23.9 39.1 27.2
T Odae, etc. - - -
K2-wx G Odae, etc. - - -
23N Aranghangchal, Baegseolchal, Baekogchal, Boramchal, Boseogchal, Boseogheugchal, Cheongbaekchal, Dongjinchal, Haepyeongchal, Hanareumchal, Hangangchal, Hangangchal1, Hwaseonchal, Jeogjinjuchal, Jinseolchal, Josaengheugchal, Nunkeunheugchal, Sangjuchal, Seolhyangchal, Sinseonchal, Unbaekchal, Unilchal 6.5 9.0 7.9
YR-wxmq A Miho, Wolbaek 12.3 13.6 12.9
G Odae, etc. - - -

* Correlation analysis between amylose content and marker SNP type

Table 6
Classification of Korean rice cultivars by Amylose content and KASP marker genotype
Table 6
Classification Cultivars amylose content K1-Wxa/Wxb K2-wx YR-wxmq
Intermediate amylose (CK) Odae (CK) 17.6 T G G
Nampyeong (CK) 20.5 T G G
Hwayeong (CK) 17.2 T G G
Dasan (CK) 17.0 T G G
Waxy Aranghangchal 7.6 T 23N G
Baegseolchal 7.4 T 23N G
Baekogchal 6.5 T 23N G
Boramchal 7.6 T 23N G
Boseogchal 8.0 T 23N G
Boseogheugchal 9.0 T 23N G
Cheongbaekchal 8.0 T 23N G
Dongjinchal 7.6 T 23N G
Haepyeongchal 7.9 T 23N G
Hanareumchal 6.8 T 23N G
Hangangchal 7.5 T 23N G
Hangangchal1 7.5 T 23N G
Hwaseonchal 8.1 H 23N G
Jeogjinjuchal 7.9 T 23N G
Jinseolchal 8.3 T 23N G
Josaengheugchal 8.4 T 23N G
Nunkeunheugchal 8.3 T 23N G
Sangjuchal 8.3 T 23N G
Seolhyangchal 8.3 T 23N G
Sinseonchal 7.8 T 23N G
Unbaekchal 8.4 T 23N G
Unilchal 8.9 T 23N G
Low amylose Baegjinju 11.2 T G G
Baegjinju1ho 11.3 T G G
Geonyang2 11.3 T G G
Miho 12.3 T G A
Wolbaek 13.6 T G A
High amylose Cheongwoo 25.4 G G G
Dodamssal 39.1 G G G
Goami 27.1 G G G
Goami2 32.0 T G G
Mimyeon 26.6 G G G
Miwoo 25.6 G G G
Mogwoo 25.3 G G G
Nokwoo 23.9 G G G
Palbangmi 26.2 G G G
Saegoami 27.1 G G G
Saemimyeon 26.6 G G G
Shingil 23.6 T G G
Yeongwoo 26.7 G G G
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Classification of Korean Rice Cultivars through Amylose Content Using Molecular Markers Targeting the Waxy Locus
Korean. J. Breed. Sci.. 2021;53(4):361-372.   Published online December 1, 2021
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Classification of Korean Rice Cultivars through Amylose Content Using Molecular Markers Targeting the Waxy Locus
Korean. J. Breed. Sci.. 2021;53(4):361-372.   Published online December 1, 2021
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Classification of Korean Rice Cultivars through Amylose Content Using Molecular Markers Targeting the Waxy Locus
Image Image Image Image
Fig. 1 Amylose content distribution of 285 Korean rice cultivars
Fig. 2 Location of SNPs of each KASP marker on Waxy gene
Fig. 3 KASP SNP marker genotyping result graph
Fig. 4 Gel-based marker genotyping result
Classification of Korean Rice Cultivars through Amylose Content Using Molecular Markers Targeting the Waxy Locus

Amylose Content and Heading Date of 285 Korean rice cultivars

Cultivars Ecotype Heading Date* Amylose Content (%)
Odae Japonica 8/3 17.6
Unbong Japonica 8/2 16.5
Geumo Japonica 8/4 17.5
Jinmi Japonica 8/6 16.6
Obong Japonica 8/2 17.7
Namwon Japonica 7/31 18.9
Jinbu Japonica 8/3 12.8
Jinbuol Japonica 7/25 16.4
Sinunbong Japonica 8/4 17.6
Sangju Japonica 8/4 17.3
Dunnae Japonica 8/3 17.8
Joryeong Japonica 8/4 18.6
Unjang Japonica 8/2 16.9
Samcheon Japonica 8/4 17.2
Junghwa Japonica 8/4 16.6
Keumobyeo2 Japonica 8/15 16.6
Guru Japonica 8/5 17.4
Sangjuchal Japonica 8/7 8.3
Undoo Japonica 7/30 16.6
Manan Japonica 8/5 18.5
Jeogjinju Japonica 8/4 17.0
Taebong Japonica 8/2 16.4
Jungsan Japonica 8/7 18.1
Saesangju Japonica 8/5 17.0
Manchoo Japonica 8/4 17.5
Manho Japonica 8/3 18.0
Namil Japonica 8/7 18.1
Joan Japonica 8/2 16.6
Heugkwang Japonica 8/13 18.3
Goun Japonica 8/1 16.6
Unkwang Japonica 8/4 17.6
Josaengheugchal Japonica 8/6 8.4
Odae1ho Japonica 8/3 17.1
Sinunbong1 Japonica 8/3 18.0
Manna Japonica 8/2 18.1
Keumo3 Japonica 8/2 18.8
Hwangkeumbora Japonica 8/4 19.0
Sandeuljinmi Japonica 8/5 18.2
Junamjosaeng Japonica 8/5 18.6
Pyeongwon Japonica 8/4 16.6
Unmi Japonica 8/4 16.9
Handeul Japonica 8/9 17.4
Jogwang Japonica 8/5 18.1
BoSeog Japonica 8/1 19.4
Joami Japonica 8/5 17.6
Hanseol Japonica 7/31 18.2
Joun Japonica 7/31 18.5
Geumyoung Japonica 8/4 17.7
Manjong Japonica 8/13 19.2
Wolbaek Japonica 8/4 13.6
Jopyeong Japonica 8/1 18.3
Jeogjinjuchal Japonica 8/7 7.9
Cheongbaekchal Japonica 8/2 8.0
Seolemi Japonica 8/5 17.5
Joeunheukmi Japonica 8/4 16.9
Saeodae Japonica 8/3 19.8
Jinseolchal Japonica 7/27 8.3
Jungmo1024 Japonica 8/4 18.0
Sanhomi Japonica 8/4 18.0
Hwawang Japonica 8/6 17.6
Nunkeunheugchal Japonica 8/6 8.3
Asemi Japonica 8/5 19.2
Ondami Japonica 8/5 18.0
jungmo1032 Japonica 7/30 18.9
Jinok Japonica 8/1 17.8
BaegIlmi Japonica 7/26 17.2
Asemi1ho Japonica 8/8 18.7
Haedamssal Japonica 8/6 19.7
Unilchal Japonica 8/7 8.9
Jungmo1043 Japonica 8/5 19.1
Jinhan Japonica 8/3 17.9
Jopum Japonica 7/30 18.0
Joil Japonica 8/6 17.6
Danpyeong Japonica 8/5 19.0
Unbaekchal Japonica 8/4 8.4
Jinkwang Japonica 8/6 17.1
Seongsan Japonica 8/7 18.6
Jonong Japonica 8/9 19.2
Shinpyeong Japonica 8/4 17.4
CW92MR Japonica 8/7 17.2
Haedeul Japonica 8/10 17.4
IS592BB Japonica 8/5 16.7
Sinseonchalbyeo Japonica 8/11 7.8
Giho Japonica 8/15 17.7
Hwaseong Japonica 8/16 17.9
Yeongdeog Japonica 8/14 18.5
Palgong Japonica 8/12 18.0
Hwajin Japonica 8/15 19.3
Janngan Japonica 8/12 18.3
Seoan Japonica 8/13 17.2
Hwayeong Japonica 8/14 17.2
Hwaseonchal Japonica 8/12 8.1
Gancheok Japonica 8/13 19.0
Hwajung Japonica 8/12 19.8
Daeripbyeo 1 Japonica 8/15 20.1
Nongan Japonica 8/16 16.6
Juan Japonica 8/13 17.3
Keumobyeo 1 Japonica 8/11 17.9
Naepung Japonica 8/10 17.4
Daejin Japonica 8/17 18.6
Sura Japonica 8/15 17.6
Hwabong Japonica 8/16 18.1
Jinpum Japonica 8/13 19.2
Sobi Japonica 8/15 17.8
Seolhyangchal Japonica 8/11 8.3
Sampyeong Japonica 8/15 18.6
Hwaan Japonica 8/14 19.4
Manpung Japonica 8/16 19.4
Haepyeong Japonica 8/12 17.2
Saegyehwa Japonica 8/18 18.9
Manwol Japonica 8/16 18.9
Yeongan Japonica 8/17 19.1
Geuman Japonica 8/18 18.5
Daepyeong Japonica 8/18 18.2
Samdeog Japonica 8/15 18.3
Pungmi Japonica 8/12 16.9
Boseogchal Japonica 8/14 8.0
Haepyeongchal Japonica 8/14 7.9
Seoan1 Japonica 8/17 19.4
Geunnun Japonica 8/17 16.5
Cheonga Japonica 8/12 18.4
Gangbaek Japonica 8/16 18.7
Haechanmulgyeol Japonica 8/14 19.2
Donghaejinmi Japonica 8/17 19.4
Cheongdam Japonica 8/15 19.3
Hongjinju Japonica 8/16 17.9
Cheongan Japonica 8/15 19.2
Borami Japonica 8/18 20.4
Heugseol Japonica 8/16 17.4
Haeoreumi Japonica 8/16 17.8
Boseogheugchal Japonica 8/16 9.0
Migwang Japonica 8/19 19.1
Cheongnam Japonica 8/15 20.1
Suryeojinmi Japonica 8/15 18.8
Gangchan Japonica 8/19 17.5
Suan Japonica 8/17 20.0
Shinbaeg Japonica 8/20 18.6
Dongbo Japonica 8/18 18.3
Jungsaenggold Japonica 8/15 16.2
Seonhyangheukmi Japonica 8/14 17.5
Daebo Japonica 8/15 17.1
Geonyang2 Japonica 8/18 11.3
Dabo Japonica 8/17 18.0
Dodamssal Japonica 8/17 39.1
Bodrami Japonica 8/17 19.6
Sinbo Japonica 8/19 18.8
Jungmo1034 Japonica 8/15 19.8
Haepum Japonica 8/18 19.2
Seonpum Japonica 8/16 17.7
Youngbo Japonica 8/15 17.6
Samkwang1 Japonica 8/16 17.2
Cheongpum Japonica 8/15 17.7
Sangbo Japonica 8/14 19.1
Heugjinmi Japonica 8/17 17.9
Boramchal Japonica 8/15 7.5
Jeogjinju2 Japonica 8/18 18.8
Dongjin Japonica 8/19 19.4
Daecheong Japonica 8/17 19.0
Tamjin Japonica 8/21 18.2
Kyehwa Japonica 8/20 17.5
Ilpum Japonica 8/18 18.2
Manguem Japonica 8/17 18.7
Hwanam Japonica 8/16 18.2
Daean Japonica 8/17 18.7
Yangjo Japonica 8/15 19.3
Hwasin Japonica 8/20 20.7
Hyangnam Japonica 8/18 18.4
Ilmi Japonica 8/18 18.7
Dongan Japonica 8/17 20.1
Hwasam Japonica 8/20 18.3
Daesan Japonica 8/21 19.4
Nampyeong Japonica 8/19 20.5
Heugnam Japonica 8/17 19.6
Aranghangchal Japonica 8/18 7.6
Hoan Japonica 8/19 19.5
Dongjinchal Japonica 8/16 7.6
Mihyang Japonica 8/17 19.5
Sindongjin Japonica 8/18 20.1
Sujin Japonica 8/21 19.1
Hojin Japonica 8/15 20.2
Heughyang Japonica 8/22 21.4
Junam Japonica 8/20 19.3
Goami Japonica 8/19 27.1
Dongin1 Japonica 8/20 19.6
Baegjinju Japonica 8/21 11.2
Seolgaeng Japonica 8/18 19.7
Goami2 Japonica 8/20 32.0
Manmi Japonica 8/18 13.6
Samkwang Japonica 8/19 19.3
Hopyung Japonica 8/19 19.5
Seopyeong Japonica - 19.0
Hwarang Japonica - 18.3
Gopum Japonica 8/17 18.8
Cheongho Japonica 8/16 20.2
Hanmaeum Japonica 8/16 18.9
Pungmi1 Japonica 8/10 16.7
Onnuri Japonica 8/17 19.3
Hanam Japonica 8/18 18.3
Dongjin2 Japonica 8/17 18.7
Baegjinju1ho Japonica 8/21 11.3
Hwangkeumnuri Japonica 8/20 19.4
Dami Japonica 8/19 19.7
Malgeumi Japonica 8/19 19.2
Hopum Japonica 8/17 18.4
Saenuri Japonica 8/19 20.2
Chilbo Japonica 8/17 18.4
Hwanggeumnodeul Japonica 8/19 19.3
Baegseolchal Japonica 8/16 7.4
Cheongcheongjinmi Japonica 8/17 20.5
Jinbaek Japonica 8/20 20.2
Dacheong Japonica 8/23 18.8
Jinsumi Japonica 8/19 19.3
Jungmo1006 Japonica 8/18 19.2
Deuraechan Japonica 8/19 19.1
Haiami Japonica 8/18 18.5
Baekogchal Japonica 8/21 6.5
Danmi Japonica 8/20 13.5
Cheonghaejinmi Japonica 8/19 19.3
Honong Japonica 8/21 19.3
Seomyeong Japonica 8/18 19.6
Younghojinmi Japonica 8/21 18.9
Jinbo Japonica 8/16 18.4
Boramchan Japonica 8/19 19.3
Anmi Japonica 8/16 16.9
Mipum Japonica 8/22 19.0
Chinnong Japonica 8/20 18.1
Geongganghongmi Japonica 8/20 20.1
Sodami Japonica 8/20 18.8
Sukwang Japonica 8/17 18.7
Huimangchan Japonica 8/16 19.0
Geonyangmi Japonica 8/18 19.7
SaeIlmi Japonica 8/19 19.7
Saegoami Japonica 8/18 27.1
Cheongun Japonica 8/18 19.4
Hyeonpum Japonica 8/20 20.5
Chindeul Japonica 8/22 17.9
Heugsujeong Japonica 8/19 18.9
Misomi Japonica 8/19 18.8
Anbaek Japonica 8/18 19.5
Manbaek Japonica 8/22 17.5
Saeshin Japonica 8/22 19.9
Cheonghyangheukmi Japonica 8/23 20.3
Sinjinbaek Japonica 8/21 19.7
Youngjin Japonica 8/20 20.3
Saechilbo Japonica 8/18 18.1
Miho Japonica 8/23 12.3
Aromi Japonica 8/20 18.1
MY299BK Japonica 8/17 18.0
Yechan Japonica 8/19 19.0
Saeilpum Japonica 8/17 17.8
Keunpum Japonica 8/15 19.3
Taebaek Tongil 8/10 17.9
Cheongcheong Tongil 8/19 17.0
Jungweon Tongil 8/12 19.3
Yongmun Tongil 8/10 16.7
Yongju Tongil 8/11 17.3
Dasan Tongil 8/12 17.0
Namcheon Tongil 8/13 17.9
Hyangmi2 Tongil 8/10 21.3
Anda Tongil 8/17 19.4
Areum Tongil 8/12 19.1
Hanareum Tongil 8/18 19.2
Dasan1 Tongil 8/14 17.3
Keunseom Tongil 8/18 17.4
Hangangchal1 Tongil 8/19 7.5
Dasan2 Tongil 8/14 18.6
Hanareum2 Tongil 8/15 17.2
Palbangmi Tongil 8/16 26.2
Mimyeon Tongil 8/13 26.6
Saemimyeon Tongil 8/20 26.6
Hanareumchal Tongil 8/19 6.8
Shingil Tongil 8/19 23.5
Hangangchal Tongil 8/19 7.5
Nampung Tongil 8/17 19.6
Jangseong Tongil 8/14 18.5
Hyangmibyeo1 Tongil 8/16 18.0
Segyejinmi Tongil 8/10 18.0
Hanareum3 Tongil 8/15 17.3
Hanareum4 Tongil 8/16 17.7
Geumgang1 Tongil 8/20 18.9
Mogwoo Tongil 8/29 25.3
Nokwoo Tongil 9/2 23.9
Yeongwoo Tongil 8/19 26.7
Cheongwoo Tongil 8/26 25.4
Miwoo Tongil 8/21 25.6

Distribution of Amylose content of 285 Korean rice cultivars

Ecotype waxy Low amylose Intermediate amylose High amylose
Japonica 19 5 223 4
Tongil 3 0 22 9

KASP marker information targeting Waxy gene

Marker Target SNP Primer Sequence
Allele X (FAM) Allele Y (HEX) Common
K1-Wxa/Wxb G/T TCATCAGGAAGAACATCTGCAAGG TCATCAGGAAGAACATCTGCAAGT GCCCAACACCTTACAGAAATTAGCA
K2-wx G/23N TTCCAGGGCCTCAAGCCCC GTTCCAGGGCCTCAAGCCCA CGCTGGTCGTCACGCTGA
YR-wxmq A/G GATGAACACACGGTCGACTCCAT ATGAACACACGGTCGACTCCAC GAGAGGGTGAGGTTTTTCCATTGCTA

Gel-based marker information targeting Waxy gene

Marker Name Chr. Sequence Annealing Temp. (℃) Restriction Enzyme Traits related Reference
Forward Reverse
Wx_a,b 6 TGTTGTTCATCAGGAAGAACATCTCCAAG TTAATTTCCAGCCCAACACC 55 EcoT14I, 37℃, 1 h~over night high amylose Yamanaka et al. 2004. Theor Appl Genet 108:1200-1204
Glu-23 6 TGCAGAGATCTTCCACAGCA GCTGGTCGTCACGCTGAG 60 waxy Wanchana et al. 2003. Plant Sci 165: 1193-1199
Wx-mq 6 GGTGAGGTTTTTCCATTGCTACAAGCA GCCCTGGTAGGAGATGTTGTGGAT 69 low amylose Sato et al. 2002. Breeding Sci 52:131-135
CAAGTACGACGCAACCACGGTAAG GGTGGACTAGACGATCTGGGTTCAA

Cultivars classified by Waxy SNP marker genotypes and their amylose content distribution

Marker SNP Cultivar Amylose Content (%)
Min. Max. Average
K1-Wxa/Wxb G Cheongwoo, Dodamssal, Goami, Mimyeon, Miwoo, Mogwoo, Nokwoo, Palbangmi, Saegoami, Saemimyeon, Yeongwoo 23.9 39.1 27.2
T Odae, etc. - - -
K2-wx G Odae, etc. - - -
23N Aranghangchal, Baegseolchal, Baekogchal, Boramchal, Boseogchal, Boseogheugchal, Cheongbaekchal, Dongjinchal, Haepyeongchal, Hanareumchal, Hangangchal, Hangangchal1, Hwaseonchal, Jeogjinjuchal, Jinseolchal, Josaengheugchal, Nunkeunheugchal, Sangjuchal, Seolhyangchal, Sinseonchal, Unbaekchal, Unilchal 6.5 9.0 7.9
YR-wxmq A Miho, Wolbaek 12.3 13.6 12.9
G Odae, etc. - - -

Classification of Korean rice cultivars by Amylose content and KASP marker genotype

Classification Cultivars amylose content K1-Wxa/Wxb K2-wx YR-wxmq
Intermediate amylose (CK) Odae (CK) 17.6 T G G
Nampyeong (CK) 20.5 T G G
Hwayeong (CK) 17.2 T G G
Dasan (CK) 17.0 T G G
Waxy Aranghangchal 7.6 T 23N G
Baegseolchal 7.4 T 23N G
Baekogchal 6.5 T 23N G
Boramchal 7.6 T 23N G
Boseogchal 8.0 T 23N G
Boseogheugchal 9.0 T 23N G
Cheongbaekchal 8.0 T 23N G
Dongjinchal 7.6 T 23N G
Haepyeongchal 7.9 T 23N G
Hanareumchal 6.8 T 23N G
Hangangchal 7.5 T 23N G
Hangangchal1 7.5 T 23N G
Hwaseonchal 8.1 H 23N G
Jeogjinjuchal 7.9 T 23N G
Jinseolchal 8.3 T 23N G
Josaengheugchal 8.4 T 23N G
Nunkeunheugchal 8.3 T 23N G
Sangjuchal 8.3 T 23N G
Seolhyangchal 8.3 T 23N G
Sinseonchal 7.8 T 23N G
Unbaekchal 8.4 T 23N G
Unilchal 8.9 T 23N G
Low amylose Baegjinju 11.2 T G G
Baegjinju1ho 11.3 T G G
Geonyang2 11.3 T G G
Miho 12.3 T G A
Wolbaek 13.6 T G A
High amylose Cheongwoo 25.4 G G G
Dodamssal 39.1 G G G
Goami 27.1 G G G
Goami2 32.0 T G G
Mimyeon 26.6 G G G
Miwoo 25.6 G G G
Mogwoo 25.3 G G G
Nokwoo 23.9 G G G
Palbangmi 26.2 G G G
Saegoami 27.1 G G G
Saemimyeon 26.6 G G G
Shingil 23.6 T G G
Yeongwoo 26.7 G G G
Table 1 Amylose Content and Heading Date of 285 Korean rice cultivars

*Transplanted on 4th June, 2020

Table 2 Distribution of Amylose content of 285 Korean rice cultivars
Table 3 KASP marker information targeting Waxy gene
Table 4 Gel-based marker information targeting Waxy gene
Table 5 Cultivars classified by Waxy SNP marker genotypes and their amylose content distribution

* Correlation analysis between amylose content and marker SNP type

Table 6 Classification of Korean rice cultivars by Amylose content and KASP marker genotype