CAJ | 학술논문

为了研究甘露糖正向筛选体系在巨尾桉遗传转化过程中的有效性，构建了以6-磷酸甘露糖异构酶（6-phosphomannose isomerase ， PMI）为筛选标记的pCAMBIA1301植物表达载体，并将该载体通过农杆菌介导的遗传转化转入木本植物巨尾桉中。将获得的阳性植株通过氯酚红（ chlorophenol red ， CPR）法及PCR检测，桉树遗传转化的阳性率达到26.09％。另外，通过正交试验优化法，对巨尾桉组培快繁体系建立过程中不同浓度激素配比进行了研究，建立起良好的巨尾桉组织培养再生体系，由甘露糖筛选敏感性测试，获得了巨尾桉筛选临界浓度，蔗糖与甘露糖比例为19∶11，优化了巨尾桉遗传转化体系，为今后巨尾桉组织培养与遗传转化研究提供了重要的参考依据。
위료연구감로당정향사선체계재거미안유전전화과정중적유효성，구건료이6-린산감로당이구매（6-phosphomannose isomerase ， PMI）위사선표기적pCAMBIA1301식물표체재체，병장해재체통과농간균개도적유전전화전입목본식물거미안중。장획득적양성식주통과록분홍（ chlorophenol red ， CPR）법급PCR검측，안수유전전화적양성솔체도26.09％。령외，통과정교시험우화법，대거미안조배쾌번체계건립과정중불동농도격소배비진행료연구，건립기량호적거미안조직배양재생체계，유감로당사선민감성측시，획득료거미안사선림계농도，자당여감로당비례위19∶11，우화료거미안유전전화체계，위금후거미안조직배양여유전전화연구제공료중요적삼고의거。
In order to study on validity of positive selection system of mannose in the transformation of Eucalyptus grandis ×E.uro-phylla, the plant expression vector pCAMBIA 1301 with 6-phosphate isomerase ( 6-phosphomannose isomerase , PMI ) as selection marker was constructed and transformed into Eucalyptus grandis ×E.urophylla by Agrobacterium-mediated genetic transformation method.The obtained positive plants were identified through chlorophenol red (CPR) method and PCR testing to show that PMI gene was really introduced into the genomes of Eucalyptus grandis ×E.urophylla, and the genetic transformation positive rate reached 26.09%.In addition , the process of establishing tissue culture system in different concentrations of hormones was studied and the tis -sue culture regenerations system and genetic transformation system was optimized by orthogonal optimization method and mannose sensi -tivity tests of Eucalyptus grandis ×E.urophylla The ratio of sucrose and mannose was 19∶11.