,2018,38(3):0456-0464 犃犮狋犪犅狅狋. 犅狅狉犲犪犾. 犗犮犮犻犱犲狀狋. 犛犻狀. :1000 4025(2018)03 0456 09 犱狅犻 :10.7606/j.issn.1000 4025.2018.03.0456 "#$%&' ( ) 犚犳 1 ()*+,, ( 6 7 2*+8, 9 750105) - %:: 49;-.&' #( ),<= > 犚犳 1? @ A B C D E? F G <H,I<= >JK@ABLM NO,: 犚犳 1@ A B, J " P Q R B S " T U VE 1WXY 1# B Z[ \]^ :(1)_ 11` #( ) 犚犳 1 犪 / 犚犳 1 犪?a,b -.& ' #( )B 22% (2) 犚犳 1 犫? @ A c \ ] _ 17 ` A B,U 2459 2283 2059 1694 2205 1784 2315@A>= #( ) B,1576 1881 1970(1985)@ A B (3) -. B,U@A, #( ) ; @A <, _ #( ) ;= #( ) _8` @A (4)NO, -. & ' #( ) : O, O < JK B-. #( ),/ O O #( ) BLM,, ` # A 9` 7`./0: &'; J"QR?;CDE ;?FG 1 + :Q789 :A 犘狅犾狔犿狅狉狆犺犻狊犿犃狀犪犾狔狊犻狊狅犳犚犳 1 犌犲狀犲犔狅犮狌狊狅犳犕犪犻狀犚犻犮犲犞犪狉犻犲狋犻犲狊 ( 犔犻狀犲狊 ) 犻狀犖犻狀犵狓犻犪 WANGJian,LIU Wei (TheCropResearchInstituteofNingxiaAcademyofAgriculturalandForestrySciences,Yongning,Ningxia750105,China) 犃犫狊狋狉犪犮狋 :49samplesselectedfrom mainvarieties(line)ofricefrom Ningxiaaretakenasexperimentalma terial,thefunctionalmarkerofwhicharesubjecttoanalysisandgenesequencecomparisonat 犚犳 1gene locusanddiscrepanciesinalmaterialsatvariouslociaresubjecttoclusteranalysistodeterminepatern andcharacteristicof 犚犳 1locusandofertheoreticalreferencefortrans breeding,molecularmarkerselec tion,configurationofhybridcombinationandutilizationofhybridvigorofsterilelineandrestoreline.the resultsshowthat:(1)11varieties (lines)are ( 犚犳 1 犪 / 犚犳 1 犪 )genetype,accountingfor22% ofmainrice variety (line)inningxia;(2) 犚犳 1 犫 genelocusdetectionfound mutationatbasicgroupof17points,a mongwhichloci2459,2283,2059,1694,2205,1784and2315havethehighestfrequencyofmutationa mongeachvariety (line),whileloci1576,1881,1970 (1985)havethelowestfrequencyofmutation;(3) Themutationsaremainlyreplacementofnucleicacidcharacterizedbynumerousloci,includinglargevaria tionamongvarieties(lines)andrelevantlessinsertionmutationlocus,onlyafewvarieties(lines).there areabout8mutationlociineachvariety (Line)inaverage;(4)Itisobservedfromclusteranalysisthat mainricevarieties(lines)inningxiacouldbedividedinto3types,witheachtypecorrespondingtomain :2017 10 21; :2018 01 17 : (31160273); (NZ14185);!"#$%&' # "% (2013NYY20305)!: (1976-),(,)*+,,-./0&'1# *+ E mai:wjzwssd@163.com 23:4 5,*+,,-./0&'1# *+ E mai:lwzws@163.com
3, : -.&' #( ) 犚犳 1?@A 457 varieties(lines)atdiferentperiods,diferenceofvarieties(lines)fromthefirstclustertothethirdcluster growsandbecomesmorediverse,butmutationofeachvarietydecreasedfrom9to7inaverage. 犓犲狔狑狅狉犱狊 :Ningxiarice;malesterilerestoregene;functionalmark;genesequence JKB,U1W 8^ B =# 1W,H,:T, 1W [1] 1# B B _ B &' ' ' _ B,> V _ LM [2],1W X Oa, B 1 #,>1W'"# _ >B [3 4]?, B XYTU, 1W&'D *+B A [5] > a 1 W '," # S " BT a 'J " [6], Q D! ' QR,?" ' #_QR? [7] $%& [8] ' Q,/ '! : BQR?, a 1 W ' C ( )*B+, -,U.? 犚犳 1 / 0 @ 1, 犚犳 1 @ A 2 ` B " Q R? 犚犳 1 犪 犚犳 1 犫?,RF1A @ RF1B ' J "/ ( 狉犳 1/ 狉犳 1) a P a Q R B Q R?( 犚犳 1/ 犚犳 1)> 犚犳 1 犪 @A >1bp 574 bp 2 3 4,> 犚犳 1 犫 @ A > 2 L M [9 10] B*+5 ^,& ' 犚犳 1 @ A > 6 _ B [11 13], &'B 78 [14],. &'9:; 'W 6 6 [15] < =,/ J K 6 > # 1 W, a?@b&'jab &' C D<> #EB-. #( ) 犚犳 1 @ A, 犚犳 1 @ A A J " P Q R BS",S"E VE,= #B Oa # B 1WXY 1# B Z[ 1 1.1 2 3 B -.;> #P> B # ( ) 49`, ^1 1.2 456789:; F 犚犳 1 犪?@A ( 狉犳 1 犪 / 狉犳 1 犪 )HQR ( 犚犳 1 犪 / 犚犳 1 犪 )3 574bpBG [9 10],>34H I J K > Rfa 7F(GGACCGGGGGATTTTAC CTG ), IJK> Rfa 7R(AACCCAACTGAGAC CATGCC) [16] F 犚犳 1 犫? F G (GenBank DQ311054.1),JK> Rfb 1F/Rfb 1R(tctcgtcgtct ctcccg/c taggcccctctgacca) Rfb 2F/Rfb 2Rc aatggacaaggccatggag/tgtaggcca tcaaacagga)< = #( )L, MNO <L G F 1.3 ' 犇犖犃 <=> 犘犆犚 :; B-. ( )# 6 7 2*+8 6,2016=4 ",5 PQ,>&'3~4 R S TRG,U CTAB S DNA > V W W )*XY_ O Y PCR 20μL,U 10 Bufer( Mg 2+ 15 mmol/l)2 μl,dntp(=2.5mmol/l)1μl,100μmol/l > =0.1μL,1U 犜犪狇,DNA Z[1μL, ddh 2O \]20μL PCR F:94 ^ 5 min; 94 30s,60 _`30s,72 ab40s, 35`c ;9 72 ab10min L E 10g/L, Gold View >,I 2 \] 2.1 "#$%&' 犚犳 1 犪?@ Rfa 7F/Rfa 7R > < < -. # ( )B DNA L,\]( 1)^,U '1 29 30 11 ` # ( )L 957bpG,( 犚犳 1 犪 / 犚犳 1 犪 )?ab 22%,U BL 383bpBG,( 狉犳 1 犪 / 狉犳 1 犪 )?ab 78% 2.2 "#$%&' 犚犳 1 犫?@ = @A > # B ^ 2, U 2459 2283 2059 1694 2205 1784 2315 @ A >= # B,= B > 39~48 1927 1117(1118) 1530(1546)@ A B@A < <,= B >11 ~21 1576 1881 1970(1985)@ A,= 1,U K A > @A 1`
458 38 A1 "#49B$%&' ( ) CDE Table1 Basicinformationof49ricevarieties(lines)inNingxia No Name Pedigree WB01 '29Jingdao29 '29Jingdao29 WB02 >39 Jingyin39 5 / 4 Fujisaka5/Hokuriku4 WB03 '216Ningdao216 6 /8023Ningjing6/8023 WB04 Akihikar / Toyonishiki/Liming WB05 '1 Ningxiangdao1 2 Jingxiangdao2 WB06 '2 Ningxiangdao2? //A30/ 76 2Xiangxuenuo//A30/Nongyuan6 2 WB07 '3 Ningxiangdao3? //A30/ 76 2Xiangxuenuo//A30/Nongyuan6 2 WB08 5 Ningnuo5 5 Ningnuo5 WB09 3 Ningjing3 >39/ >59Jingyin39/Jingyin59 WB10 7 Ningjing7 12/66 6// / J Hongqi12/66 6/Liming//Shanfengbuzhi WB11 9 Ningjing9 78 4442/78 127 WB12 12 Ningjing12 80K 479 1/ 15280K 479 1/Qingza52 WB13 13 Ningjing13 3 /IR24//76 1303Keqing3/IR24//76 1303 WB14 14 Ningjing14 8131F1/8035F1 WB15 15 Ningjing15 87F 129/84XZ 7 WB16 16 Ningjing16 81D 86/81K 249 3 WB17 18 Ningjing18 777 61/ 69 7Nongyuan77 61/Ji69 7 WB18 19 Ningjing19 83XW 555/86JZ 12 WB19 23 Ningjing23 88XW 495 1/84XZ 7 WB20 24 Ningjing24 12 / 4 //92 37Ningjing12/Italia4//92Xiawen37 WB21 25 Ningjing25 10 / 76 2Ningjing10/Nongyuan6 2 WB22 26 Ningjing26 >2 Jinyin2 WB23 27 Ningjing27 11 2/ Leng11 2/Saturn WB24 28 Ningjing28 >1 / 21Shanyin1/Hua21 WB25 29 Ningjing29 >1 /91XW 65Shanyin1/91XW 65 WB26 30 Ningjing30 31116S+30301S+5047S+4018S/ 1 + 2 31116S+30301S+5047S+4018S/Chaochan1+Chaochan2 WB27 31 Ningjing31 LQ 1 WB28 32 Ningjing32 83XZ 489/ 12583XZ 489/Tenxi125 WB29 33 Ningjing33 93JZ 5/93H 1 (1) WB30 34 Ningjing34 552A/Q15552A/Hui15 WB31 35 Ningjing35 88xw 495 1/84xz 7 WB32 36 Ningjing36 6/// 259/02428// 8503/84xZ 7You6///Zhonzuo59/02428//Zhongxi8503/84xZ 7 WB33 37 Ningjing37 96G 59/Dongnong41596G 59/ 415 WB34 38 Ningjing38 /96G 59Jingfen/96 59 WB35 40 Ningjing40 /96G 59Jingfen/96 59 WB36 41 Ningjing41 94xw 127/X 11 94xw 127/Zupei11 WB37 42 Ningjing42 89XW 216/92XW 723 WB38 43 Ningjing43 12 / 4 //92 37Ningjing12/Italia4//92Xiawen37 WB39 44 Ningjing44 C57 80 9(101209)/C79 6 WB40 45 Ningjing45 152/ 9502Jiuxin152/Kai9502 WB41 46 Ningjing46 12 / 4 //92 37Ningjing12/Italia4//92Xiawen37 WB42 47 Ningjing47 '1 / 101 Wuyoudao1/Jijing101 WB43 98XW 258 98XW 258 WB44 51 Hua51 51 Hua51 WB45 114 Hua114 114 Hua114 WB46 843Nongke843 843Nongke843 WB47 105Jijing105 2 / 89 45Chaochan2/Ji89 45 WB48 784Jing784 784Jing784 WB49 28 Chujing28 26 /96Y 6Chujing26/96Y 6
3, : -.&' #( ) 犚犳 1?@A 459 WB01~WB49.K^1;M.DL2000 1 49;-.&' #( ) 犚犳 1 犪?@ACDE WB01 WB49areshowninTable1;M:DL2000 Fig.1 Functionalmarkanalysisofmainricevarieties(lines)in49ricesamplesat 犚犳 1 犪 genelocus A2 Table2 "#49B$%&' ( ) Numberofvarietiesateachmutationpointofmain ricevarieties(lines)in49ricesamples @ Mutationposition Quantity @ Mutationposition Quantity 1117(1118) 19 1927 21 1233 11 1970(1985) 1 1530(1546) 14 2059 47 1576 1 2205 46 1694 47 2283 47 1784 46 2315 39 1881 1 2459 48 E > Rfb 1F/Rfb 1R Rfb 2F/Rfb 2R < < # ( )DNA L I F,\ ] 犚犳 1 犫?FG<H,_17`AB (^ 3), > 1117 @ A G T,1118 @ A C T,1233@A C T,1530@A C T,1546@ A G A,1576@A A AC,1694@A C T,1784 @ A C T,1881 @ A T TC, 1927@A C A,1970@A A AG,1985@ A A AG,2059@A T G,2205@A T C,2283@A C G,2315@A A G,2459 @A G C U 1117 1118 @ A,1530 1546@A,1970 1985@A^ K,> K E U K A = # @ A ^ 4 ^ 4, _1` #_10 ` @A,22 ` #_9` @A,U B # 9 ` @A,13 ` #_8` @A,6 ` #_7` @A,4 ` #_6` @A,2` #_5` @A, : ( @AB # 5, 5`B @A #_ 32 ` #BFG 犚犳 1 犫? 2.3 "#$%&' 犚犳 1 +, DPS, FK ;B # B Nei Li,U O <49 ;&' NO,I NO,\] 28 : 1.25, : O, O_ 43 24 12 13 ` # > 1117(1118) 1694 1784 1927 2059 2205 2283 2315 2459@ A,> 1233 1530(1546) 1576 1881 1970 (1985)@A, @A P O _ 28 843 27 11 ` # > 1233 1694 1784 2059 2205 2283 2315 2459 @ A # ( ),> 1117 (1118) 1576 1927 1970(1985)@A8_ #( ),1530(1546)@ A # ( ), 1881@A _ #( ) # B LM-.>1881 1530(1546)@ O _ 105 44 ' 29 25 ` # _1233 1881 #,U @ B _, # BLM
460 38 3 49 ( ) 犚犳 1 犫 Table3 犚犳 1 犫 genemutationlociof49ricevarieties(lines)ningxia Mutationlocusposition 1117(1118) 1233 1530(1546) 1576 1694 1784 1881 1927 1970(1985) 2059 2205 2283 2315 2459 WB01 - - C T,G A - C T C T - - - T G T C C G A G G C WB02 G T,C T - - - C T C T - C A - T G T C C G A G G C WB03 - C T C T,G A - C T C T - - - T G T C C G A G G C WB04 G T,C T - - - C T C T - C A - T G T C C G A G G C WB05 - - - - C T C T - - - T G T C C G A G G C WB06 G T,C T - - - C T C T - C A - T G T C C G A G G C WB07 G T,C T - - - C T C T - C A - T G T C C G A G G C WB08 - C T C T,G A - C T C T - - - T G T C C G A G G C WB09 G T,C T - - - C T C T - C A - T G T C C G A G G C WB10 G T,C T - - - C T C T - C A - T G T C C G A G G C WB11 - - C T,G A - - - - - - T G - C G A G G C WB12 G T,C T - - - C T C T - C A - T G T C C G A G G C WB13 G T,C T - - - C T C T - C A - T G T C C G A G G C WB14 G T,C T - - - C T C T - C A - T G T C C G A G G C WB15 G T,C T - - - C T C T - C A - T G T C C G A G G C WB16 G T,C T - - - C T C T - C A - T G T C C G A G G C WB17 - C T C T,G A - C T C T - - - T G T C C G A G G C WB18 - C T C T,G A - C T C T - - - T G T C C G A G G C WB19 - C T - - C T C T - - - T G T C C G A G G C WB20 G T,C T - - - C T C T - C A - T G T C C G A G G C WB21 - C T C T,G A - C T C T - - - T G T C C G A G G C WB22 - - C T,G A - C T C T - - - T G T C C G A G G C WB23 - C T - - C T C T - - - T G T C C G A G G C WB24 - C T C T,G A - C T C T - - - T G T C C G A G G C WB25 - - - - C T C T - C A - T G T C C G - G C WB26 - - - - C T C T - - - T G T C C G - G C
3, : -.&' #( ) 犚犳 1?@A 461 3 ContinuedTable3 Mutationlocusposition 1117(1118) 1233 1530(1546) 1576 1694 1784 1881 1927 1970(1985) 2059 2205 2283 2315 2459 WB27 - C T C T,G A - C T C T T TC - - T G T C C G A G G C WB28 - - - - - - - - - - - - - - WB29 G T,C T - - - C T C T - C A - T G T C C G A G G C WB30 G T,C T - - - C T C T - - - T G T C C G - G C WB31 - - - - C T C T - C A - T G T C C G A G G C WB32 - - C T,G A - C T C T - - - T G T C C G A G G C WB33 - - - - C T C T - C A - T G T C C G - G C WB34 - - C T,G A - C T C T - - - T G T C C G A G G C WB35 - - - - C T C T - - - T G T C C G - G C WB36 - - - A AC C T C T - - A AG,A AG, T G T C C G - G C WB37 - - - - C T C T - - - T G T C C G A G G C WB38 G T,C T - - - C T C T - C A - T G T C C G A G G C WB39 G T,C T - - - C T C T - C A - T G T C C G A G G C WB40 - - - - C T C T - - - T G T C - A G G C WB41 - - C T,G A - C T C T - - - T G T C C G A G G C WB42 - - C T,G A - C T C T - - - T G T C C G A G G C WB43 - - - - C T - - - - T G T C C G - G C WB44 G T,C T - - - C T C T - C A - - - C G A G G C WB45 G T,C T - - - C T C T - C A - T G T C C G - G C WB46 - C T - - C T C T - - - T G T C C G A G G C WB47 G T,C T - - - C T C T - C A - T G T C C G A G G C WB48 - - - - C T C T - - - T G T C C G - G C WB49 - C T - - C T C T - - - T G T C C G A G G C
462 38 Table4 A4 "#49B$%&' ( ) Numberofmutationlociofeachvarietyof49ricevarieties(lines)inNingxia @A Numberof mutationlocus @A Numberof mutationlocus @A Numberof mutationlocus @A Numberof mutationlocus WB01 8 WB14 9 WB27 10 WB39 9 WB02 9 WB15 9 WB28 0 WB40 6 WB03 9 WB16 9 WB29 9 WB41 8 WB04 9 WB17 9 WB30 7 WB42 8 WB05 7 WB18 9 WB31 8 WB43 5 WB06 9 WB19 8 WB32 8 WB44 7 WB07 9 WB20 9 WB33 7 WB45 8 WB08 9 WB21 9 WB34 8 WB46 8 WB09 9 WB22 8 WB35 6 WB47 9 WB10 9 WB23 8 WB36 8 WB48 6 WB11 5 WB24 9 WB37 7 WB49 8 WB12 9 WB25 7 WB38 9 WB13 9 WB26 6 WB01~WB49K^1 2 49;-.&'B 犚犳 1?@ANO WB1 WB49areshowninTable1 Fig.2 Treediagramfor 犚犳 1genelocusclusteranalysisof49ricevarietiesinNingxia 3?X B-. [17 18], B ', JKB,> WB K ' - ' [19 21],? ' > V B L M [22 24] E < B-. 犚犳 1 犫? F G < H,\ ] _ 2B@A, ` @A B JK BOa-. B, @A, # B @A <, _ #( ), D B [25] F 犚犳 1? J K @ A, : -.B # 3`O, O : " B #( ), O "B # ( ), O H OB, AB O B # B # ( ), @ A, A O O, # @ABLM,, AB _ < - D JK B E _ : "# E-.,"#E B P ( D & JAB, #J.., <
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