过表达LrAN2基因对马铃薯中花青素和糖苷生物碱含量的影响  

Effects of overexpression of LrAN2 gene on contents of anthocyanins and glycoalkaloids in potato

作　　者：李红艳 李洁雅 李响 叶广继[1,2,3,4,5,6] 周云 王舰[1,2,3,4,5,6] LI Hong-Yan;LI Jie-Ya;LI Xiang;YE Guang-Ji;ZHOU Yun;WANG Jian(Qinghai University,Xining 810016,Qinghai,China;Qinghai Academy of Agriculture and Forestry Sciences,Xining 810016,Qinghai,China;Qinghai University Key Laboratory of Qinghai-Tibet Plateau Biotechnology of the Ministry of Education,Xining 810016,Qinghai,China;Qinghai Provincial Key Laboratory of Potato Breeding,Xining 810016,Qinghai,China;Provincial and Ministry Co-construction of the State Key Laboratory of Sanjiangyuan Ecology and Plateau Agriculture and Animal Husbandry,Xining 810016,Qinghai,China;Laboratory for Research and Utilization of Qinghai Tibet Plateau Germplasm Resources,Xining 810016,Qinghai,China)

机构地区：[1]青海大学,青海西宁810016 [2]青海省农林科学院,青海西宁810016 [3]青海大学青藏高原生物技术教育部重点实验室,青海西宁810016 [4]青海省马铃薯育种重点实验室,青海西宁810016 [5]省部共建三江源生态与高原农牧业国家重点实验室,青海西宁810016 [6]青藏高原种质资源研究与利用实验室,青海西宁810016

出　　处：《作物学报》2023年第4期988-995,共8页Acta Agronomica Sinica

基　　金：财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-9);青海省科学自然基金项目-创新团队(2022-ZJ-902);青海省创新平台建设专项(2022-ZJ-Y01)资助。

摘　　要：目前对马铃薯(Solanum tuberosum L.)中MYB基因参与的花青素研究较为深入,但对影响马铃薯品质和安全的糖苷生物碱(steroidal glycoalkaloids,SGAs)在不同组织中的变化规律及调控机制尚不完全清楚。LrAN2是黑果枸杞(Lycium ruthenicum)中的MYB基因,与黑果枸杞果实中花青素的积累有关。本研究以野生型大西洋、2种转LrAN2大西洋株系(LrAN2oe#66和LrAN2oe#200)为试验材料,检测不同组织中的花青素和SGAs含量,并对SGAs生物合成相关基因的表达量分析。pH示差法检测花青素发现,仅在转基因植株(LrAN2oe#200)叶片中检测到一定量的花青素(12 mg 100 g-1FW)。高效液相色谱-串联质谱法检测SGAs发现,3个材料不同组织SGAs含量变化为叶片>薯皮>薯肉。薯皮中的SGAs含量无显著性差异;薯肉中LrAN2oe#66的SGAs含量较对照降低、LrAN2oe#200对照显著增加1.3倍,但未超出安全标准(0.2mg g-1FW);叶片中LrAN2oe#66的SGAs含量较对照增加1倍、LrAN2oe#200较对照显著增加3.8倍。qRT-PCR分析基因表达量发现,StHMG1和StSGT2基因受到LrAN2基因的调控在转基因植株中显著上调。本研究结果对马铃薯植株中花青素含量的积累具有指导意义,为进一步解析马铃薯资源中花青素及SGAs的调控机制提供理论依据。At present,the research on the anthocyanins involved in the MYB gene in potato(Solanum tuberosum L.)is relatively in-depth,but the change rule and regulatory mechanism of the glycoalkaloids(steroidal glycoalkaloids,SGAs)in different tissues,which affect the quality and safety of potato,are still unclear.Lr AN2,the MYB gene in Lycium ruthenicum,is associated with the accumulation of anthocyanins in Lycium ruthenicum fruit.In this study,to detect the contents of anthocyanins and SGAs in different tissues,and to analyze the expression of genes related to SGA biosynthesis,wild-type Atlantic and two trans-LrAN2 Atlantic lines(LrAN2oe#66 and LrAN2oe#200)were used as the test materials.The detection of anthocyanins by pH differential method showed that only a certain number of anthocyanins(12 mg 100 g-1FW)were detected in the leaves of transgenic plants(LrAN2oe#200).The contents of SGAs by HPLC-TMS in different tissues of the three materials were as follows:leaves>potato skin>potato flesh.There was no significant difference in the contents of SGAs in potato peels.Compared with the control,the SGAs content of LrAN2oe#66 in potato flesh was lower than the control,and the SGAs content of LrAN2oe#200 was significantly increased by 1.3 times,but did not exceed the safety standard(0.2 mg g-1FW).The contents of SGAs in LrAN2oe#66 of leaves were increased by 1 times and LrAN2oe#200 was significantly increased by 3.8 times compared with the control.StHMG1 and StSGT2 genes were significantly up-regulated in transgenic plants under the regulation of LrAN2 gene by qRT-PCR.These results give a guiding significance for the accumulation of anthocyanins in potato plants and provide a theoretical basis for further analysis of the regulation mechanism of anthocyanins and SGAs in potato resources.

关 键 词：LrAN2基因 马铃薯 花青素 糖苷生物碱 

分 类 号：S532[农业科学—作物学]