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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (4): 988-995.doi: 10.3724/SP.J.1006.2023.24082

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

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

LI Hong-Yan1(), LI Jie-Ya1, LI Xiang1, YE Guang-Ji1,2,3,4,5,6, ZHOU Yun1,2,3,4,5,6, WANG Jian1,2,3,4,5,6,*()   

  1. 1Qinghai University, Xining 810016, Qinghai, China
    2Qinghai Academy of Agriculture and Forestry Sciences, Xining 810016, Qinghai, China
    3Qinghai University Key Laboratory of Qinghai-Tibet Plateau Biotechnology of the Ministry of Education, Xining 810016, Qinghai, China
    4Qinghai Provincial Key Laboratory of Potato Breeding, Xining 810016, Qinghai, China
    5Provincial and Ministry Co-construction of the State Key Laboratory of Sanjiangyuan Ecology and Plateau Agriculture and Animal Husbandry, Xining 810016, Qinghai, China
    6Laboratory for Research and Utilization of Qinghai Tibet Plateau Germplasm Resources, Xining 810016, Qinghai, China
  • Received:2022-04-03 Accepted:2022-07-21 Online:2023-04-12 Published:2022-08-17
  • Contact: *E-mail: jianwang2197@163.com
  • Supported by:
    China Agriculture Research System of MOF and MARA(CARS-9);Qinghai Natural Science Foundation Program-Innovation Team(2022-ZJ-902);Qinghai Innovation Platform Construction Project(2022-ZJ-Y01)

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Abstract:

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. LrAN2, 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-1 FW) 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-1 FW). 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.

Key words: LrAN2 gene, potato, anthocyanin, glycoalkaloid

Fig. 1

Potato SGA biosynthesis pathway"

Table 1

Primer sequences used in this study"

引物名称
Primer name		 正向引物
Forward sequence (5'-3')		 反向引物
Reverse sequence (5'-3')
qStHMG1 CAGGTTCAAATGCAAGACTC GATAGATCTATTATATTTCAT
qStSQS1 ACTTGCAGAGACTCGGGAAC TCGGTTGCCAGAAAGTTGTG
qStCAS ACCATTACACTCTGCAGCAA GAACGATATTCTCCCAATGC
qStSSR2 AAGCGCCTTGAACAGAGGAA ACCCTTGACATTTGGCCCAT
qStGAME4 GGCTTGCATTTGAGGTGTTTA GAGCCTTGAGTCCCTTATGAT
qStGAME8a TGGGAGATATGACAATTC GCCATGCCAAAGTTATTACC
qStSGT1 GGAACAATCTCACTGCTC CACACACACACCAAGTTAC
qStSGT2 CAATCTTACCGCACTTATAG GTGTTTATTCCCAGCCCTAG
qStSMT1 GATATCTAGGGGACAGGT CAGGTAGCTTCTATTGCG
qStCYP51G AATGTGGGGTTGCTGTTAGT ATCAACCCACCAACAATAGG
Actin (internal reference gene) AGATGCTTACGCTGGATG GAATGC TTCCGGTGTGGTTGGATTCTGTTC

Fig. 2

Partially overexpressed LrAN2 gene in potato M: DL 2000 marker; Plasmid: LrAN2 plasmid; WT: wild-type."

Fig. 3

Phenotypic characteristics of Atlantic and transgenic potatoes (A), anthocyanin content of Atlantic, and LrAN2oe#200 (B) n.d. means the anthocyanin content is 0 mg 100 g-1, biological replicates n = 3."

Fig. 4

SGAs content in potato peel (A), flesh (B), and leaf (C) in Atlantic, LrAN2oe#66, and LrAN2oe#200 Different lowercase letters indicate significant difference in the contents of SGAs at P < 0.05."

Fig. 5

Relative expression levels in SGAs biosynthesis-related genes in Atlantic and LrAN2oe#200 potato peels Different lowercase letters indicate significant differences in the relative expression levels among different varieties at P < 0.05."

Fig. 6

Relative expression levels in SGAs biosynthesis related genes in Atlantic and LrAN2oe#200 fleshes Different lowercase letters indicate significant differences in gene expression among different varieties at P < 0.05."

Fig. 7

Relative expression levels of SGAs biosynthesis-related genes in the leaves of Atlantic and LrAN2oe#200 Different lowercase letters indicate significant differences in gene expression among different varieties at P < 0.05."

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