| 龙葵UNUSUAL FLORAL ORGANS类SnUFO2基因C端序列的保守性对花发育的影响 |
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| 引用本文： | 周佳圆,钟玉,努尔阿斯娅·伊马木,崔敏龙,朴春兰.龙葵UNUSUAL FLORAL ORGANS类SnUFO2基因C端序列的保守性对花发育的影响[J].浙江农林大学学报,2023,40(1):38-44. |
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| 作者姓名： | 周佳圆 钟玉 努尔阿斯娅·伊马木 崔敏龙 朴春兰 |
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| 作者单位： | 1.浙江农林大学 园艺科学学院，浙江 杭州 3113002.乌什县农业农村局，新疆 阿克苏 843006 |
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| 基金项目： | 宁夏回族自治区重点研发计划重大项目(2019BFG02011) |
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| 摘 要： | 目的 UNUSUAL FLORAL ORGANS (UFO)基因属于F-box基因家族，是重要的花器官特征基因。UFO基因N端能与Skp1类基因结合形成Skp1-Cullin1-F-box (SCF)复合体，参与泛素化过程并降解C端结合的靶蛋白。为了探究C端序列对龙葵Solanum nigrum花发育的影响，本研究克隆了一个C末端缺失的SnUFO2*基因并构建其表达载体转入龙葵植株中，观察转基因龙葵植株花器官变化，从而深入探讨UFO基因完整的C末端序列在龙葵花发育中的重要作用。 方法 利用生物信息学分析软件对SnUFO2*和全长的SnUFO2比较分析，采用实时荧光定量PCR(RT-qPCR)对SnUFO2*基因在野生型龙葵植株根、茎、叶、花苞中进行表达分析；通过超表达载体的构建、转基因植株表型的观察及石蜡切片技术验证SnUFO2基因的功能。 结果 SnUFO2*基因ORF长度为1302 bp，编码433个氨基酸，与龙葵中完整的SnUFO2基因相比，其C末端缺失了23个氨基酸。RT-qPCR结果显示:SnUFO2*基因在野生型植株的花苞中特异性表达。对转基因植株的表型观察发现:35S:: SnUFO2*转基因龙葵植株的花瓣向萼片转化。石蜡切片分析发现:转基因龙葵植株雄蕊缺失，雌蕊处有不确定的分生组织产生。 结论 35S:: SnUFO2*转基因龙葵植株花瓣、雄蕊和心皮发育异常。C端结构缺失可能降低了SnUFO2蛋白特异性识别靶蛋白的能力，说明该基因完整的C末端对龙葵花器官发育至关重要。图5表1参23
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| 关 键 词： | 转基因龙葵 SnUFO2 花分生组织 花器官发育 花发育 |
| 收稿时间： | 2022-03-16 |
Effect of conservation of C-terminal sequence of Solanum nigrum UNUSUAL FLORAL ORGANS family SnUFO2 on flower development |
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| ZHOU Jiayuan,ZHONG Yu,Nurasiya Imam,CUI Minlong,PIAO Chunlan.Effect of conservation of C-terminal sequence of Solanum nigrum UNUSUAL FLORAL ORGANS family SnUFO2 on flower development[J].Journal of Zhejiang A&F University,2023,40(1):38-44. |
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| Authors: | ZHOU Jiayuan ZHONG Yu Nurasiya Imam CUI Minlong PIAO Chunlan |
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| Institution: | 1.College of Horticulture Science, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China2.Wushi County Bureau of Agriculture and Rural Affairs, Aksu 843006, Xinjiang, China |
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| Abstract: | Objective UNUSUAL FLORAL ORGANS (UFO) gene belongs to the F-box gene family and is one of the most important floral organ identity genes. The N-terminal of UFO gene can combine with Skp1 genes to form Skp1-Cullin1-F-box (SCF) complex, which participates in the ubiquitination process and degrades the target protein binding with the C-terminal. In order to explore the effect of C-terminal sequence on the flower development of Solanum nigrum, a C-terminal deleted SnUFO2* gene was cloned and its expression vector was constructed transferred into S. nigrum plants to observe the changes of floral organs of transgenic S. nigrum plants, so as to further explore the important role of the complete C-terminal sequence of UFO gene in the flower development of S. nigrum. Method Comparative analysis of SnUFO2* and full-length SnUFO2 was performed using bioinformatics analysis software. Moreover, the expression levels of SnUFO2* in roots, stems, leaves, and buds of wild-type S. nigrum were analyzed by real-time quantitative PCR (RT-qPCR); the function of the SnUFO2* gene was verified by the construction of over expression vector, observation of phenotype of transgenic plants, and the employment of paraffin section technique. Result The ORF length of SnUFO2* was 1 302 bp, encoding 433 amino acids, when compared with the complete SnUFO2 in S. nigrum, the C-terminal of which was missing 23 amino acids. RT-qPCR results showed that SnUFO2* was specifically expressed in the buds of wild-type plants. The phenotypic observation of transgenic plants demonstrated that the petals of 35S::SnUFO2* transgenic S. nigrum plants transformed into sepals. Paraffin section analysis presented that the stamens of transgenic S. nigrum plants were missing and uncertain meristem was produced in the pistil. Conclusion 35S::SnUFO2* transgenic plants lead to abnormal development of petals, stamens, and carpels of S. nigrum. The deletion of the C-terminal structure may reduce the ability of SnUFO2 protein to specifically recognize target proteins, indicating that the complete C-terminal of the gene is important for organ development of S. nigrum. Ch, 5 fig. 1 tab. 23 ref. ] |
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