运用ISSR分子标记鉴定杉木 × 侧柏远交杂种

 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。图3表1参17

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

To identify truthfulness of distant hybrids for Cunninghamia lanceolata（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） × B1 distant cross combinations of C. lanceolata × 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；polyacrylamide 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.］