葡萄种质离体保存中低温和矿物油覆盖延长继代间隔的效应

Efficacy of Low Temperature and Mineral Oil Covering in Prolonging Subculture Interval During Grape Germplasm Preservation in Vitro

以‘巨峰’等14个葡萄品种试管苗为试材,研究不同保存温度和培养基表面覆盖矿物油对其生长的影响。结果表明:低温可有效延长试管苗的继代间隔时间,转入常温培养后恢复生长良好,但不同品种适宜的温度不同。设定存活率降至30%~40%时的保存时间为最长继代时间,‘红斯威特’‘莫丽莎’‘皇家夏天’‘克瑞森无核’葡萄最适低温9℃,可保存850 d以上,部分可至1060 d;‘巨峰’‘巨玫瑰’‘皇家秋天’‘火焰无核’‘夏黑’‘公主’‘粒粒特’‘赤霞珠’‘美人指’‘魏可’最适低温12~15℃,可保存350~600 d。葡萄试管苗低温保存前需常规培养一段时间(预培养),并根据培养温度及品种决定适宜的预培养时间,一般为7~21 d。接种不带叶片的单芽茎段并立即在培养基表面覆盖6~8 cm厚矿物油能将多个葡萄品种试管苗在常温下的继代间隔时间由150 d延长至420 d,将矿物油倒出,试管苗即可恢复生长。

Fourteen grape cultivars were used as plant materials to reveal the influences of low culture temperature and mineral oil covering on medium of plantlets in vitro.The results showed that the low temperature could significantly prolong the subculture period,but the optimal low temperature varied according to the different cultivars.Plantlets cultured in the low temperature could resume growth well after being transferred to routine temperature culture.When the survival rate of 30%–40%was a concern,the optimal temperature was 9℃for’Sweet Scarlet’,’Melissa’,’Summer Royal’and’Crimson Seedless’cultivars,and the subculture period could reach more than 850 days,and maximum 1060 days for certain cultivar;the optimal temperature was 12–15℃for’Kyoho’,’Jumeigui’,’Autumn Royal’,’Flame Seedless’,’Summer Black’,’Princess’,’Lilit’,’Cabernet Sauvignon’,’Manicure Finger’and’Wink’,and the subculture period could last 350 to 600 days.Prior to the low temperature culture,the plantlets needed to be cultured under routine condition for a period of time(preculture).The duration depended on the culture temperature and the varieties,and it was usually 7 to 21 days.By inoculating shoots with single bud and without leaf,as well as adding 6–8 cm mineral oil on the top of the medium,the subculture period could be up to 420 days in routine temperature culture condition while the control was 150 days.The plantlets resumed growth after the mineral oil was removed.