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作物学报 ›› 2025, Vol. 51 ›› Issue (4): 982-991.doi: 10.3724/SP.J.1006.2025.44147

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

I型MADS-box基因SlMADS79调控番茄株型的功能研究

郭绪虎1,*(), 李灵芝2, 李凤1, 马博岩1, 贾晓宇1   

  1. 1山西大同大学农学与生命科学学院, 山西大同 037009
    2山西农业大学园艺学院, 山西晋中 030801
  • 收稿日期:2024-09-08 接受日期:2024-12-12 出版日期:2025-04-12 网络出版日期:2024-12-12
  • 通讯作者: 郭绪虎, E-mail: 429248441@qq.com
  • 基金资助:
    山西省基础研究计划项目(202303021211181);大同市应用基础研究计划项目(2024067);山西大同大学基础研究项目(2022Q3)

Functional study on the regulation of plant architecture by tomato type I MADS-box gene SlMADS79

GUO Xu-Hu1,*(), LI Ling-Zhi2, LI Feng1, MA Bo-Yan1, JIA Xiao-Yu1   

  1. 1School of Agronomy and Life Sciences, Shanxi Datong University, Datong 037009, Shanxi, China
    2College of Horticulture, Shanxi Agricultural University, Jinzhong 030801, Shanxi, China
  • Received:2024-09-08 Accepted:2024-12-12 Published:2025-04-12 Published online:2024-12-12
  • Contact: E-mail: 429248441@qq.com
  • Supported by:
    Basic Research Programs of Shanxi Province(202303021211181);Applied Basic Research Program of Datong City(2024067);Basic Research Programs of Shanxi Datong University(2022Q3)

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摘要：

MADS-box基因在植物生长发育全过程中具有重要调控功能。目前, 关于番茄II型MADS-box基因的功能研究较多, 而I型MADS-box基因却鲜有报道。本研究克隆了I型MADS-box基因SlMADS79, 该基因在番茄根、叶和侧芽中表达水平相对较高, 推测其可能参与番茄营养器官生长调控。本研究以经典番茄栽培品种Ailsa Craig (AC++)为背景材料, 采用RNA干扰技术沉默SlMADS79基因。与野生型相比, SlMADS79基因沉默株系顶端优势度降低、植株矮小; 叶片的长度、宽度、周长以及面积均小于野生型植株; 根系总长度、总表面积、总投影面积、根系体积、分叉数、根尖数均显著减少。解剖学研究表明, 沉默植株茎秆的纵切面细胞较小, 然而其平均数量明显增加。在激素水平上, SlMADS79沉默株系中IAA (吲哚-3-乙酸)、TZR (转玉米素核糖苷)以及CS (栗木甾酮)含量降低。在分子水平上, 生长素应答基因IAA3和赤霉素合成基因GA3ox1在SlMADS79沉默株系中显著下调, 而细胞周期基因CyCA3;1在沉默株系中显著上调。本研究从形态、解剖、激素和分子水平上进一步解析了SlMADS79基因的生物学功能, 丰富了番茄I型MADS-box基因家族的研究, 为番茄株型调控研究提供了可靠的理论依据。

关键词: 番茄, MADS-box, SlMADS79, 株高, 叶片形态, 根系

Abstract:

MADS-box genes play a crucial role in regulating plant growth and development. While the functions of type II MADS-box genes have been extensively studied, there are relatively few reports on type I MADS-box genes in tomato. In this study, we cloned the type I MADS-box gene SlMADS79, which was found to be highly expressed in tomato roots, leaves, and lateral buds, suggesting its involvement in the regulation of vegetative organ growth. Using the classical tomato cultivar Ailsa Craig (AC++) as background material, we silenced the SlMADS79 gene through RNA interference (RNAi). Compared to the wild type, SlMADS79-silenced lines exhibited reduced apical dominance and decreased plant height. The length, width, perimeter, and area of the leaves were smaller than those of wild-type plants. Additionally, root traits—including total length, total surface area, total projected area, volume, number of forks, and number of tips—were significantly reduced. Anatomical studies revealed that while the cells in the longitudinal sections of SlMADS79-silenced stems were smaller, their average number significantly increased. At the hormonal level, the contents of IAA (indole-3-acetic acid), TZR (trans-zeatin riboside), and CS (castasterone) were decreased in the SlMADS79-silenced lines. At the molecular level, the auxin response gene IAA3 and gibberellin synthesis gene GA3ox1 were significantly downregulated in the SlMADS79-silenced lines, while the cell cycle gene CyCA3;1 was significantly upregulated. This study further analyzes the biological function of the SlMADS79 gene at morphological, anatomical, hormonal, and molecular levels, expands our understanding of the type I MADS-box gene family in tomato, and provides a solid theoretical foundation for further research on plant architecture regulation in tomato.

Key words: tomato, MADS-box, SlMADS79, plant height, leaf morphologies, root system

表1

本研究所用引物"

引物名称Primer name 引物序列Primer sequences (5'-3') 引物用途Primer usage
Forward fragment-F ATGGTGAGAGGGAAAACTGAAATG 植物表达载体构建
Plant expression vector construction
Forward fragment-R CCTTGAGGTTGAAAATTCATAAAGTTTGTC
Reverse fragment-F CCTTGAGGTTGAAAATTCATAAAGTTTGTC
Reverse fragment-R ATGGTGAGAGGGAAAACTGAAATG
trans-MADS79-F AGAAGACGTTCCAACCACG 转基因植株鉴定
Transgenic plant identification
trans-MADS79-R CCAACTTGAGCATCACAAAGAAC
SlCAC-F CCTCCGTTGTGATGTAACTGG 内参引物
Internal control primer
SlCAC-R ATTGGTGGAAAGTAACATCATCG
SlMADS79-F GAAATGAGGCGTATCGAAAA qRT-PCR检测
qRT-PCR detection
SlMADS79-R TCCAACTTGAGCATCACAAA
IAA3-F GGCCACCAGTTCGATCATAC
IAA3-R GGTGCTCCATCCATGCTAAC
GA3ox1-F GTAGACCAAAGGAACCCTCAAAT
GA3ox1-R GCCGAACAGATGAAAGTGCT
CycA3;1-F CTAAGAAAAGAGCAGCAGAAGCA
CycA3;1-R GATTCCTTATCTTTTTCAGCAACAG
DELLA-F CAGATTCATCAGCAACGAGACC
DELLA-R TGTGAAACCGCAAGAATACCAA
GAI-F CCAGCACTTGTCATTCTTACCC
GAI-R AAAGCTCATCCATTCCAGCA

图1

SlMADS79基因在野生型番茄中的表达模式分析 基于qPCR的SlMADS79在野生型番茄不同发育阶段中的相对表达模式。RT: 根; ST: 茎; YL: 幼叶; ML: 成熟叶; SL: 衰老叶; BR: 分枝; F: 花; IMG: 未成熟的青果实; MG: 成熟的青果实; B: 破色期的果实; B4: 破色后4 d的果实; B7: 破色后7 d的果实。"

图2

SlMADS79基因RNAi载体(A)与转基因植株PCR验证(B) M: DL2000; 10~20为T0代株系; WT为阴性对照(野生型); CK为空白对照(水); P为阳性对照(农杆菌菌液)。"

图3

SlMADS79基因沉默植株的表型 A: 野生型植株幼苗形态; B, C: 转基因株系幼苗形态; D: 野生型和转基因植株高度测量。*表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

图4

野生型和SlMADS79转基因株系茎秆的解剖分析 A: 野生型番茄茎的纵切面; B: 转基因番茄茎的纵切面。标尺为100 μm; C: 估测的野生型和转基因株系茎秆的细胞面积; D: 估测的野生型和转基因株系茎秆的细胞周长; E: 估测的野生型和转基因株系茎的细胞数量。*表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

图5

野生型和SlMADS79转基因株系叶片形态 A: 野生型(黄色下画线)和转基因株系(红色下画线)叶片照片; B~E: 野生型和SlMADS79转基因系叶片的长度、宽度、周长和面积。*表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

图6

野生型和SlMADS79转基因株系根系形态 A: 野生型(黄色下画线)和转基因株系(红色下画线)根系照片; B~G: 野生型和SlMADS79转基因株系根系的总长度、总表面积、总投影面积、根系体积、分叉数、根尖数。*表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

图7

野生型和SlMADS79沉默株系茎秆的激素水平 *表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

图8

野生型和SlMADS79沉默植株中植物激素响应、信号转导以及细胞周期调控相关基因的转录水平 *表示野生型和转基因株系之间存在显著差异(P < 0.05)。"

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