运用ISSR分子标记鉴定杉木×侧柏远交杂种

Hybrid identification of Cunninghamia lanceolata × Platycladus orientalis based on ISSR markers

为了对杉木Cunninghamia lanceolata与侧柏Platycladus orientalis远交杂种的真实性进行鉴定,进行预备试验,从100条UBC primer Set#9(800～900)引物中,筛选出9条多态性引物。对杉木×侧柏远交组合A4(1419)×B1所得杂交后代中选择的4株超级苗等材料,提取DNA,进行聚合酶链式反应(PCR)扩增,并按母本、远交子代和父本顺序,对PCR产物进行电泳实验。结果发现,4株超级苗在7条引物中,共找到16条父系特征谱带,在另外2条引物中检测到父母的共同谱带。由此可以肯定,杂交所得超级苗是杉木与侧柏远缘杂交产生的杂种。杉木与侧柏远缘杂交是可配的,杉木远缘杂交育种是一条多性状改良途径。

To identify truthfulness of distant hybrids for Cunninghamia huweolata （Chinese fir） × Platycladus orientalis, a series of exploratory experiments were conducted with 9 intersimple sequence repeat （ISSR） primers having polymorphism screened out from 100 University of British Columbia （UBC） primer Set # 9 （800 - 900）. For A4 （1419） × BI distant cross combinations of C. laneeolata x P. orientalis, four outstanding seedlings were selected. Analysis of the parents and the four outstanding seedlings was done with DNA extraction; polymerase chain reaction （PCR） amplification using 9 polymorphism primers; polyaery lamide gel electrophoresis（PAGE） based on the following order of maternal ： A4, seedlings （No. 2, 3, 11, 12）, paternal, and ethylene dibromide （EB） staining; and DNA band observations with an ultraviolet lamp. Then, these were recorded with an FR-200A Bio-electrophoresis Image Analysis System. Experimental results of the four outstanding seedlings （No. 2, 3, 11, 12） showed that for the combination of A4 × B1 ： （1） there were 16 paternal special bands in 7 of the 9 polymorphic primers and （2） there were no paternal special bands with the other 2 primers, but some common bands of their parents were detected within the four seedlings. Thus, （1） all selected outstanding seedlings were distant hybrids; （2） distant hybridization between C. lanceolata × P. orientalis was compatible; and （3） wide cross breeding of Chinese fir could be used for multiple-trait genetic improvement of forest trees. [Ch, 3 fig. 1 tab. 17 ref.]