桑树MaNAC90的克隆及其在干旱胁迫下的功能分析

doi:10.16420/j.issn.0513-353x.2025-0214

摘要/Abstract

摘要：

以‘德果1号’桑树（Morus alba‘Deguo 1’）为材料，克隆了1个NAC转录因子基因MaNAC90，其开放阅读框为732 bp，编码243个氨基酸。MaNAC90蛋白分子量为27.436 kD，等电点为5.32，含有1个NAC保守结构域，与TERN亚族成员聚为一类，定位在细胞核和细胞膜。模拟干旱胁迫（200 mmol · L^-1甘露醇）下，与野生型相比，过表达MaNAC90的拟南芥的发芽率显著降低，根长显著变短，幼苗的萎蔫程度更严重，MDA含量和电导率均显著升高，RD22和RD29B的表达水平显著下调。本研究结果初步表明MaNAC90在干旱胁迫响应中发挥负调控作用。

关键词: 桑, 果用, MaNAC90, 干旱胁迫, 功能分析

Abstract:

A NAC transcription factor gene MaNAC90 was cloned from Morus alba‘Deguo 1’. The open reading frame of MaNAC90 was 732 bp，encoding 243 amino acids with molecular weight 27.436 kD and isoelectric point 5.32. MaNAC90 contained one NAC domain and belonged to TERN group，was localized in the nucleus and the cell membrane. Under simulated drought stress（200 mmol · L^-1mannitol），transgenic Arabidopsis expressing MaNAC90 exhibited a significant reduction in germination rate，a notable decrease in root length，an increased severity of seedling wilting，and significantly increased MDA content and electrolyte leakage，along with significantly decreased expression of RD22 and RD29B when compared to wild-type plants. These results preliminary indicated that MaNAC90 plays a negative regulatory role in response to drought stress.

Key words: mulberry, fruit use, MaNAC90, drought stress, function analysis

黄仁维, 李晓冬, 邓红梅, 杨晏, 曾睿, 杨财容. 桑树MaNAC90的克隆及其在干旱胁迫下的功能分析[J]. 园艺学报, 2026, 53(5): 1274-1284.

HUANG Renwei, LI Xiaodong, DENG Hongmei, YANG Yan, ZENG Rui, YANG Cairong. Cloning and Functional Analysis of Mulberry MaNAC90 Gene Under Drought Stress[J]. Acta Horticulturae Sinica, 2026, 53(5): 1274-1284.

图/表 9

图1 MaNAC90基因的PCR扩增 1：MaNAC90条带；2：阴性对照

Fig. 1 PCR amplification of MaNAC90 gene 1：MaNAC90 fragment；2：Negative control

图2 MaNAC90蛋白与其他植物NAC的多序列比对 Ma：桑树；Me：木薯（XP_021629180.1）；Mr：杨梅（KAB1200809.1）；Cs：大麻（XP_030503399.2）；Hb：橡胶树（XP_021678053.1）；Pa：糙叶山麻黄（PON65421.1）。A ~ E：亚结构域

Fig. 2 Multiple sequence alignment of MaNAC90 with NAC protein other plants Ma：Morus alba；Me：Manihot esculenta（XP_021629180.1）；Mr：Morella rubra（KAB1200809.1）；Cs：Cannabis sativa（XP_030503399.2）；Hb：Hevea brasiliensis（XP_021678053.1）；Pa：Parasponia andersonii（PON65421.1）. A-E：Subdomain

图3 MaNAC90与拟南芥NAC蛋白的进化树分析

Fig. 3 Phylogenetic tree analysis of MaNAC90 and NAC proteins from Arabidopsis

图4 MaNAC90蛋白的亚细胞定位

Fig. 4 Subcellular localization of MaNAC90 protein

图5 MaNAC90转基因拟南芥DNA鉴定（A）及表达量检测（B） WT：野生型；OE：过表达株系。下同

Fig. 5 DNA identification（A）and expression level analysis（B）of MaNAC90 transgenic Arabidopsis WT：Wild type；OE：Over expression line. The same below

图6 200 mmol · L-1甘露醇处理下野生型和转基因拟南芥种子的发芽率及根长 A：发芽表型；B：发芽率；C：根长表型；D：根长。* 表示与野生型有显著差异（P < 0.05）。下同

Fig. 6 Seeds germination rate and root length of wild-type and transgenic Arabidopsis under 200 mmol · L-1 mannitol treatment A：Germination phenotype；B：Germination rate；C：Root length phenotype；D：Root length. * indicates significant differences from wild type（P < 0.05）. The same below

图7 自然干旱处理下野生型和转基因拟南芥的表型

Fig. 7 Phenotype of wild-type and transgenic Arabidopsis under natural drought treatment

图8 干旱处理下野生型和转基因拟南芥的生理指标

Fig. 8 Physiological indicators of wild-type and transgenic Arabidopsis under drought treatment

图9 干旱处理下野生型和转基因拟南芥中RD22和RD29B基因的表达水平

Fig. 9 The expression analysis of RD22 and RD29B of wild-type and transgenic Arabidopsis under drought treatment

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