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JIPB | 南京农业大学吴泽/滕年军团队揭示LlSAPK2增强百合耐热性的分子机制

  • 2026-06-24 06:58:18
JIPB | 南京农业大学吴泽/滕年军团队揭示LlSAPK2增强百合耐热性的分子机制

百合是重要的观赏花卉和切花作物，其生长、开花和商品品质对温度变化十分敏感。种球繁育和设施切花生产常跨越高温季节，短时热浪也可能造成难以恢复的叶片灼伤、花期缩短和商品率下降。全球气候变暖使极端高温事件更频繁，容易造成百合叶片萎蔫、花器官损伤、活性氧过量积累和细胞膜脂质过氧化，进而降低观赏价值与种球生产效率。植物可通过热激转录因子及其下游保护基因重建细胞稳态，SnRK2家族丝氨酸/苏氨酸蛋白激酶则负责整合多种非生物胁迫信号。然而，已有研究主要关注SnRK2在干旱、盐和低温响应中的作用，其是否直接参与园艺植物的热胁迫感知，以及磷酸化怎样快速维持关键转录因子的稳定性，仍缺少清晰证据。
2026年6月19日，南京农业大学园艺学院吴泽副教授团队和滕年军教授团队在知名期刊《JIPB》发表了题为“LlSAPK2 confers thermotolerance in lily by phosphorylating and stabilizing LlbZIP46”的研究论文。研究从百合(Lilium longiflorum )中鉴定到热诱导SnRK2家族激酶LlSAPK2，结合基因沉默、稳定遗传转化、蛋白互作、磷酸化、蛋白降解和启动子结合分析，发现热胁迫迅速激活LlSAPK2；活化的LlSAPK2通过磷酸化并稳定LlbZIP46，增强其对LlHSFA3B启动子的结合和转录激活，最终促进热响应及活性氧清除。研究还利用百合花瓣盘的瞬时遗传组合验证二者的上下游关系，将生理表型、蛋白修饰和靶基因转录串联起来，形成一个依靠转录因子快速稳定化来增强百合耐热性的调控模块，也为观赏百合耐热种质改良提供了可利用的激酶和转录因子靶点。
研究通过系统发育和序列分析发现，LlSAPK2与菠萝AcSAPK2亲缘关系最近，并与拟南芥SnRK2.7、SnRK2.8共同归入SnRK2亚类II。LlSAPK2转录在高温下快速诱导，特异抗体和以MBP为底物的激酶检测进一步显示其酶活也被迅速激活。TRV介导的LlSAPK2沉默在室温下对相对离子渗漏率和丙二醛影响较小，但在热胁迫下显著提高两项指标，并导致DAB、NBT染色显示的活性氧积累增加，同时LlHSFA1、LlHSFA2、LlHSFA3A和LlHSFA3B表达下降，说明该激酶参与细胞膜保护和抗氧化调节。
研究获得3个稳定遗传的LlSAPK2过表达株系，其转录水平达到野生型的2.8至4.5倍，蛋白积累还通过特异抗体和根尖GFP信号得到验证。过表达植株在高温下萎蔫和卷曲减轻，相对离子渗漏率、丙二醛和活性氧水平降低，4个A类HSF表达升高，基础及获得性耐热性均增强，形成与沉默材料方向相反的遗传证据。获得性耐热结果还说明，LlSAPK2不仅参与初次高温响应，也能帮助植株在预先感受热信号后建立更强的后续保护。酵母双杂交筛选得到OsbZIP46样蛋白LlbZIP46，进一步通过双分子荧光互补、荧光素酶互补和GST pull-down证明LlSAPK2与LlbZIP46可在细胞核内及体外直接互作。体内外磷酸化实验表明LlSAPK2直接磷酸化LlbZIP46；沉默LlSAPK2后，LlbZIP46在室温和高温下的Phos-tag迁移信号均减弱，说明这种修饰依赖LlSAPK2。
图1 LlSAPK2 与 LlbZIP46 相互作用以调节百合的耐热性
LC-MS/MS共鉴定到10个潜在磷酸化位点，研究者从中选择8个高可信位点，构建包含S19、S22、S43、T55的A1突变体和包含S87、S95、T98、S118的A2突变体。两组突变均削弱磷酸化，而A2区域突变对LlbZIP46稳定性的影响更明显。无细胞和体内降解实验显示，LlbZIP46在室温下较快降解，高温则明显延缓其降解；LlSAPK2过表达或与LlbZIP46共同表达都会进一步提高蛋白稳定性。这种差异使LlbZIP46在平时维持较低水平，而在高温时快速积累，兼顾正常发育和应急防御。结合CHX和MG132处理结果，研究推测磷酸化可能通过减少泛素连接酶识别、增强伴侣蛋白作用或改变蛋白构象来抵抗降解。
百合花瓣盘瞬时系统同时组合LlSAPK2沉默和LlbZIP46过表达后发现，LlbZIP46可部分挽救LlSAPK2沉默引起的严重褪色、膜损伤和活性氧积累，且LlHSFA1、LlHSFA2和LlHSFA3B的诱导对LlSAPK2依赖更明显，支持LlbZIP46是LlSAPK2下游的重要执行因子。启动子分析在LlHSFA1、LlHSFA3A和LlHSFA3B中发现bZIP基序，但酵母单杂交只验证了LlbZIP46与LlHSFA3B启动子结合。EMSA进一步定位到ACGTG元件，双荧光素酶实验显示LlbZIP46可激活LlHSFA3B启动子，共表达LlSAPK2后激活能力显著增强；增加LlSAPK2蛋白还会提高LlbZIP46对启动子的结合。
研究提出了一个以蛋白稳定性为核心的百合快速耐热模型：常温下LlSAPK2活性较低，LlbZIP46维持较低水平，避免持续启动热响应干扰正常发育；高温来临后，LlSAPK2被快速激活并磷酸化LlbZIP46，减少其降解，增强其在细胞核中的积累以及对LlHSFA3B启动子的结合，进而提高A类热激转录因子表达和活性氧清除能力。这种机制无需等待大量新蛋白合成，便可通过稳定既有调控因子迅速建立防御，实现即时保护与长期转录重编程的衔接。研究填补了SnRK2参与百合高温响应的机制空白，也为耐热百合分子育种实践提供了潜在靶点。
原文链接：https://doi.org/10.1111/jipb.70326
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  1. CONNECT:[ UseTime:0.000425s ] mysql:host=127.0.0.1;port=3306;dbname=b460;charset=utf8mb4
  2. SHOW FULL COLUMNS FROM `fenlei` [ RunTime:0.000565s ]
  3. SELECT * FROM `fenlei` WHERE `fid` = 0 [ RunTime:0.000676s ]
  4. SELECT * FROM `fenlei` WHERE `fid` = 63 [ RunTime:0.000349s ]
  5. SHOW FULL COLUMNS FROM `set` [ RunTime:0.000503s ]
  6. SELECT * FROM `set` [ RunTime:0.000212s ]
  7. SHOW FULL COLUMNS FROM `article` [ RunTime:0.000602s ]
  8. SELECT * FROM `article` WHERE `id` = 590809 LIMIT 1 [ RunTime:0.001848s ]
  9. UPDATE `article` SET `lasttime` = 1782323580 WHERE `id` = 590809 [ RunTime:0.001079s ]
  10. SELECT * FROM `fenlei` WHERE `id` = 65 LIMIT 1 [ RunTime:0.000300s ]
  11. SELECT * FROM `article` WHERE `id` < 590809 ORDER BY `id` DESC LIMIT 1 [ RunTime:0.000442s ]
  12. SELECT * FROM `article` WHERE `id` > 590809 ORDER BY `id` ASC LIMIT 1 [ RunTime:0.000897s ]
  13. SELECT * FROM `article` WHERE `id` < 590809 ORDER BY `id` DESC LIMIT 10 [ RunTime:0.000692s ]
  14. SELECT * FROM `article` WHERE `id` < 590809 ORDER BY `id` DESC LIMIT 10,10 [ RunTime:0.000700s ]
  15. SELECT * FROM `article` WHERE `id` < 590809 ORDER BY `id` DESC LIMIT 20,10 [ RunTime:0.001260s ]
0.097522s