DNA2ITS, Iden tif ica tion of C o lle to trichum sp. from A n thu rium and raeanum and Its Sequenc ing of R ibosoma l D NA2ITS

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1 2009, 36 (5) : Acta Horticulturae Sinica DNA2ITS 1, 2, 2, 2, 13 ( 1, ; 2, ) : DNA2ITS, :, : ; ; ; DNA2ITS : S : A : X (2009) Iden tif ica tion of C o lle to trichum sp. from A n thu rium and raeanum and Its Sequenc ing of R ibosoma l D NA2ITS X ING Hong2mei 1, 2, D ING Ping 2, WANG Ke2rong 2, and ZHOU Xiao2yun 13 ( 1 Guangzhou Flow er Research Center, Guangzhou , China; 2 College of Plant Protection, N anjing A gricultural U niversity, N anjing , China) Abstract: Among the Colletotrichum strains isolated from A n thu rium and raeanum jing, Yangzhou, there were two group s exhibited different morphology on the culture. in Guangzhou, Nan2 Two strains of each group were chose for species identification w ith the morphological character, pathogencity on p lants, range and the sequence of ribosomal DNA 2ITS. Colletotrichum g loeosporioides and C. and raeanum. Both species could infect solely, crossly and latently. host The results indicated that these strains belong to two species, destructivum, and the latter was first reported as a pathogen in A n thu rium Key words: A n thu rium and raeanum ; Colletotrichum ; pathogen identification; ribosom al DNA 2ITS, (Anthu rium andraeanum ) (Colletotrichum sp. ), :,,,, ;,,,, ,, 136 : ; : : (2004Z32E0411) 3 Author for correspondence ( E2mail: xyzh28@1261com; Tel: )

2 744 36, C01, HZ21, 112 YZ21 Kochp s ( ) ( ml - 1 ) 25, RH 90% 100%, 5 #,,, 8, 3, 2 d, 1 2 d,,, 113 PDA 7 d,, 25, 12 h : (Spathiphyllum kochii Engler & Krause. ) ( ), (D ieffenbachia picta) ( ), ( Spa thiphyllum floribundum L ind. et. Andre NE. B r. cv. M ) ( ), (Guzm ania continental) ( ), (Calathea m akoyana) ( ), (D racaena cam bodiana) ( ), (D endrobium sp. ) ( ), ( Phalaenopsis am abilis) ( ), (Cym bidium sp. ) (), (D racaena arborea) ( ), (M alus pum ila M ill. ) ( ), 25, RH 90% 100%, 6, ( ) 115 rd NA ( in terna l tran scr ibed space, d, 1 2 d ITS) PCR PDA PDA ( 30 ml ), 3, d,, 5 ml, h,, DNA 012 g115 ml Eppendorf, 900 L CTAB, 90 L 10% SDS,, 55 1 h, r m in m in, ( ),, r m in m in, r m in m in, 1 /10 NaAc 2, h, r m in m in, 70% 1, 30 L ITS DNA ITS1 ITS4 PCR, ITS1: 5 2TCCGTAG2 GTGAACCTGCGG23, ITS4: 5 2TCCTCCGCTTATTGATATGC23

3 5 : DNA2ITS PCR PCR 25 L: DNA, 015 mol, 4 dntp 50 mol, 215 L 10 PCR, 2 mmolmg 2 +, 1125 Taq ( Promega), 1, PE2400 PCR: 94 5 m in;, 94 1 m in, s, 72 1 m in, 31 ; 72 7 m in UN IQ210 PCR, PMD182T, E1coli JM109, Amp LBPCR, ITS DNA2ITS GenBank ( http: //www1ncbi1nlm1gov1blast) 136, PDA 2 (C038 C049; C08 C014) : ; ; C08 ( 6215% ) 212 C038 C049 PDA :, ;,,,,,, cm,,, m m,, ( 1),,, Colletotrichum gloeosporioides ( Penz. ) Penz. & Sacc. F ig. 1 C038 A: ; B: ; C: 1 M oda lity of stra in C038 in Colletotrichum sp. A: Colony; B: Conidia; C: App ressoria. C08 C014 PDA,,,,,,, ( ),, m m, C038

4 C049,,,,, ( 2), Colletotrichum destructivum OπGara F ig. 2 C014 A: ; B: ; C: 2 M oda lity of stra in C014 in Colletotrichum sp. A: Colony; B: Conidia; C: App ressoria. 213 C038 C049, 3 d,,,,, 4 d 012 cm,,, 30 d, ( 3) C08 C014 5 d, C038, C038 ( 3) :,, F ig d 3 Sym ptom on A n thu rium andraeanum leaves caused by Colletotrichum sp. 30 days after inoculation 214,,, C038 C049 :,,,,

5 5 : DNA2ITS 747 C08 C014 ; C038, 215 ITS : C038 C049 (C. g loeosprioides), C08 C014 HZ215 (C. destructivum ), DNA2ITS : C038 C049 ITS C08 C014 HZ215 85% 87% C038, C049DNA2ITS, :, DNA2ITS532 bp 534 bp B last GenBank (NCB I), 100, 98 C. gloeosporioides Glom erella cingulata, 97% 98%, 98% C. gloeosporioides ITS, 3 C08 C014 HZ215, :, 3 DNA2ITS 538 bp B last GenBank, 100, 98C. destructivum, 3 99%, C. destructivum ITS 3 ITS,,, ITS, DNA ITS ( internal transcribed space), ( Cooke et al., 1996; A ltomare et al., 1997; Johnson & Jones, 1997; Leclerc2Potvin et al., 1999;, 2000;, 2004;, 2007) (2006) ITS, (M ycosphaerella fijiensis) Yan (1995) ITS Phialophora am ericana P. verrucose, ITS, 97% ( Sreenivasp rasad et al., 1996 ), ITSC038 C049 C08 C014 HZ215,, ITS, ITS, B last GenBank AR3750 ( Colletotrihum sp. ) AR3749 ( Glom erella sp. ) ITS 99%,, C. destructivum,, ITS, (, 2008), ITS References A ltomare C, Petrini O, Logrieco A Taxonom ic relationship s among the toxigenic species Fusarium sporotrichioides and Fusarium tricinctum by isozyme analysis and RAPD assay. Can J Bot, 75 (4) :

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