双色万代兰种子形态及无菌萌发研究OA

Taking V.bicolor mature seeds refrigerated at-25℃ for 135 days as test materials,the seed morphological characteristics and viability were investigated using stereomicroscopy and 0.5％TTC staining,respectively.An orthogonal experiment was conducted using MS as the basic medium to study the effects of different combinations of 6-BA,NAA,and coconut water(CW)under light and dark conditions for 180 days,to establish a suitable asymbiotic germination system,in order to provide a reference for the germplasm conservation and artificial breeding of the endangered Vanda bicolor.The results showed that the seeds of V.bicolor were brownish-black.The seed length and width were(0.301±0.019)mm and(0.098±0.009)mm,respectively,while the embryo length and width were(0.228±0.031)mm and(0.096±0.013)mm,respectively.The embryo occupied almost the entire seed coat cavity.TTC testing indicated that the seeds maintained a viability of 60.3％after long-term low-temperature storage.In the asymbiotic germination experiment,the optimal medium combination was MS+1.0 mg·L-1 6-BA+0 mg·L-1 NAA+100 mL·L-1 CW.Light conditions significantly improved the germination rate,while dark culture was conducive to protocorm expansion and differentiation.Range analysis revealed that the primary factors affecting germination were in the order of 6-BA＞CW＞NAA.In conclusion,6-BA and CW were the key factors promoting the seed germination of V.bicolor.A phased configuration of light and dark culture was beneficial for improving germination and protocorm growth.