The Effect of Stu-miR156 Silencing by STTM Technology on Potato Lateral Root Development

doi:10.16420/j.issn.0513-353x.2020-0230

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (3): 531-538.doi: 10.16420/j.issn.0513-353x.2020-0230

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The Effect of Stu-miR156 Silencing by STTM Technology on Potato Lateral Root Development

LUO Hongyu¹, YANG Jiangwei¹, FENG Ya¹, ZHANG Huanhuan¹, LIU Shengyan¹, ZHANG Ning¹^,²^,^*(), SI Huaijun¹^,²^,^*

¹College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China
²Gansu Provincial Key Laboratory of Aridland Crop Science,Lanzhou 730070,China

Received:2020-05-10 Online:2021-03-25 Published:2021-04-02
Contact: ZHANG Ning,SI Huaijun E-mail:ningzh@gsau.edu.cn

Abstract

Abstract:

To elucidate the biological function of Stu-miR156 in potato lateral root development,plantlets in vitro of potato cultivar‘Desiree’were used as dornors. Using STTM（Short tandem target mimic,STTM）technology,the Stu-miR156 silencing expression vector was successfully constructed and further the transgenic plants were obtained viaAgrobacterium-mediated method. qRT-PCR was used to determine the differential expression of Stu-miR156 and its target gene StSPL9in the transgenic potato plants. We constructed a pCAM-GFP-StSPL9 fusion protein expression vector and transformed it into tobacco,thereby confirming that StSPL9 was localized in the cytoplasm and nucleus. The result of qRT-PCR showed that the expression levels of Stu-miR156 in roots,stems and leaves were severely inhibited,while the target gene StSPL9 was up-regulated in the transgenic potato lines L1 and L2. The number of lateral roots of the transgenic plants was significantly reduced and the growth is inhibited compared with the wild type plants.

Key words: potato, Stu-miR156, STTM, StSPL9, genetic transformation, lateral root

CLC Number:

S532

LUO Hongyu, YANG Jiangwei, FENG Ya, ZHANG Huanhuan, LIU Shengyan, ZHANG Ning, SI Huaijun. The Effect of Stu-miR156 Silencing by STTM Technology on Potato Lateral Root Development[J]. Acta Horticulturae Sinica, 2021, 48(3): 531-538.

Figures/Tables 8

Table 1 Nucleotide sequences

序列名称 Sequence name	序列（5′-3′） Sequence
Stu-miR156	UUGACAGAAGAUAGAGAGCAC
Linker（48 nt）	gttgttgttgttatggtctaatttaaatatggtctaaagaagaagaat
Stu-miR156STTM	GGATCCGTGCTCTCTCctaTCTTCTGTCAAgttgttgttgttatggtctaatttaaatatggtctaaagaagaagaatGTGCTCTCTCctaTCTTCTGTCAAAAGCTT

Fig. 1 Prediction of Stu-miR156 target gene and analysis of target gene structure

Fig. 2 Subcellular localization of the pCAM-GFP-SPL9 protein

Fig. 3 Schematic diagram of Stu-miR156STTM vector（A）and recombinant expression vector pCAMBIA1301-Stu-miR156STTM（B）

Fig. 4 PCR identification of Stu-miR156STTM transgenic plants

Fig. 5 Analysis of relative expression of the transgenic plants（L1 and L2）with Stu-miR156STTM transformation Different lowercase letters indicate significant differences（P < 0.05）.

Table 2 Phenotypic assay of the transgenic potato plants L1 and L2

株系 Plant line	株高/mm Height	根数 Root number	根长/mm Root length	根冠比/% Root-shoot ratio
野生型Wild type	72.08 ± 5.05 a	33.33 ± 3.68 a	27.33 ± 1.25 a	38.03 ± 2.03 a
L1	69.42 ± 3.37 a	5.67 ± 2.05 b	26.33 ± 1.70 ab	38.02 ± 3.09 a
L2	65.02 ± 6.51 a	8.33 ± 1.70 b	22.67 ± 1.70 b	34.97 ± 0.84 a

Fig. 6 Phenotypes of the transgenic potatoes with Stu-miR156STTM transformation

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The Effect of Stu-miR156 Silencing by STTM Technology on Potato Lateral Root Development

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