摘要
【背景】氨基酸是植物体内非常重要的有机氮化合物,氨基酸转运蛋白参与了植物中氨基酸的转运,对植物的生长发育起着重要作用。【方法】为深入研究烟草氨基酸通透酶(Amino acid permease,AAPs)基因家族成员AAP3的功能,从普通烟草红花大金元中克隆NtAAP3基因,检测其在普通烟草不同发育时期和器官中的表达差异;创制该基因敲除纯合突变体,统计突变体大田农艺性状,检测突变体中氨基酸、蛋白质和色素含量,分析突变体在正常光照和强光胁迫条件下的光合指数变化。【结果】NtAAP3基因编码区(CDS)全长为1440 bp,在叶片中高表达,尤其是旺长期和打顶期;与对照组相比,NtAAP3基因敲除纯合突变体的自然株高和打顶株高有所降低,最大腰叶长/宽无明显变化;烤后烟叶的总蛋白质含量降低,其中脯氨酸和天冬酰胺的含量下降尤为显著;叶绿素a/b、总叶绿素和类胡萝卜素含量均有所升高;正常光照条件下的最大光化学效率(Fv/Fm)值无明显变化、非光化学淬灭(NPQ)值有所上升,而强光胁迫条件下的Fv/Fm和NPQ值均显著下降。【结论】NtAAP3基因突变导致烟草中氨基酸转运和蛋白质积累的效率降低,烟叶的色素含量、光合效率以及总产量未受影响,为烟草相关性状的品种改良提供了新的研究方向。
[Background]Amino acids are crucial organic nitrogen compounds in plants,and amino acid transporters are involved in their transport,playing a significant role in plant growth and development.[Methods]To further investigate the function of AAP3,a member of the tobacco amino acid permease(AAPs)gene family,the NtAAP3 gene was cloned from Nicotiana tabacum cv.Honghua Dajinyuan.Its expression levels were examined at different developmental stages and in various organs of N.tabacum.Homozygous knockout mutants of the gene were created.The agronomic traits of the mutants were recorded in the field,and the contents of amino acids,proteins,and pigments in the mutants were measured.Additionally,the photosynthetic parameters of the mutants were analyzed under both normal and high-light stress conditions.[Results]The full-length coding sequence(CDS)of the NtAAP3 gene is 1440 bp and is highly expressed in leaves,particularly during the vigorous growing and topping stages.Compared to the control group,the homozygous NtAAP3 knockout mutants exhibited a reduction in natural plant height and post-topping plant height,while the length/width of the largest middle leaves showed no significant change.In the cured tobacco leaves,the total protein content was lower,with a particularly significant decrease in the contents of proline and asparagine.The contents of chlorophyll a/b,total chlorophyll,and carotenoids were all increased.Under normal light conditions,there was no significant change in the maximum photochemical efficiency(Fv/Fm)value,but the non-photochemical quenching(NPQ)value increased.However,under high-light stress,both the Fv/Fm and NPQ values decreased significantly.[Conclusion]The mutation of the NtAAP3 gene leads to reduced efficiency in amino acid transport and protein accumulation in tobacco.The pigment content,photosynthetic efficiency,and total yield of the tobacco leaves were not affected,providing a new research direction for the improvement of tobacco-related traits.
作者
魏攀
WEI Pan(Zhengzhou Tobacco Research Institute of CNTC,Zhengzhou 450001,China)
出处
《中国烟草学报》
北大核心
2025年第5期96-107,共12页
Acta Tabacaria Sinica
基金
豫中烟区低氯高钾新品系试验示范与工业应用(D2020186)
中国烟草总公司烟草基因组计划重大专项项目“低烟气苯酚释放量的云烟87、K326、红花大金元定向改良”[110201901019(JY-06)]。
关键词
氨基酸转运
氨基酸通透酶
实时荧光定量PCR
基因编辑
光合指数
普通烟草
amino acid transport
amino acid permease
real-time quantitative PCR
gene editing
photosynthetic parameters
Nicotiana tabacum
