烟草腺毛生物反应器材料的创制与性状分析

Creation and characterization of tobacco glandular trichome bioreactor material

为提高烟草腺毛分泌物含量(质量分数),以高分泌型种质资源T.I.1068为材料,采用基因编辑技术敲除其腺毛发育负调控基因NtCycB2,获得了纯合突变株系TTB-1(tobacco trichome bioreactor-1)。结果表明:TTB-1植株生长旺盛,发育正常。显微观察发现与烤烟K326不同,TTB-1与T.I.1068表皮毛由腺毛组成,无保护毛;TTB-1腺毛密度比T.I.1068提高53.41%,是K326的7倍;与K326相比,TTB-1腺头更加饱满,罗丹明着色更深。烟株现蕾期中部叶片叶面化学成分检测结果显示,TTB-1与T.I.1068分泌物组分一致,但分泌物含量显著高于T.I.1068,分泌物总量比T.I.1068提高63.11%;TTB-1与K326分泌物组分不同,除西柏烷类二萜和Ⅰ~Ⅱ型蔗糖酯外,还含有赖百当类二萜和Ⅲ~Ⅴ型蔗糖酯,总量是烤烟品种K326的7.95倍。对上中下部成熟叶片进行分批采收和叶面化学成分萃取,浸膏产率分别为烟叶鲜质量的0.61%、0.43%、0.30%,平均每株浸膏产量为8.90 g;在盛花期进行一次性采收,浸膏产率为烟叶鲜质量的0.45%,平均每株浸膏产量为8.06 g。

To improve the secretion mass fraction of tobacco glandular trichomes,a homozygous mutant plant TTB-1(tobacco trichome bioreactor-1)was obtained by using gene editing technology to remove NtCycB2,a negative regulatory gene of glandular trichome development,in T.I.1068 germplasm.The results showed that TTB-1 plants grew vigorously and developed normally.Unlike the flue-cured tobacco cv.K326,the surface fur of TTB-1 and T.I.1068 was composed of glandular trichomes without protection.The glandular trichome density of TTB-1 was 53.41%higher than that of T.I.1068 and 7 times higher than that of K326.Compared to K326,the TTB-1 glandular head was fuller and Rhodamine staining was darker.The test results of the chemical components on the leaf surface of the middle leaves of tobacco plants at the bud stage indicated that the components in TTB-1 and T.I.1068 secretions were consistent,but the content of secretions was significantly higher in TTB-1,and its total secretion amount was 63.11%higher than that of T.I.1068.The secretions of TTB-1 differed from those of K326.In addition to cembranoid diterpene and typeⅠ-Ⅱsucrose esters,TTB-1 also contained labdane diterpene and typeⅢ-Ⅴsucrose esters,and its total secretion amount was 7.95 times of that of K326.The mature upper,middle and lower leaves were harvested in batches and the chemical components on the leaf surfaces were extracted.The concrete yield was 0.61%,0.43%,and 0.30%of the fresh upper,middle,and lower leaf weights,respectively,and the average concrete yield per plant was 8.90 g.The concrete yield was 0.45%of the fresh leaf weight,and the average concrete yield per plant was 8.06 g when harvesting by one-priming at flowering stage.