Biotechnology Bulletin ›› 2026, Vol. 42 ›› Issue (10): 1-12.doi: 10.13560/j.cnki.biotech.bull.1985.2025-1380
ZHANG Wan-yu, XING Jun-qiang, SUN Qin-fu, WANG You-ping(
)
Received:2025-12-17
Online:2026-10-26
Published:2026-10-15
Contact:
WANG You-ping
E-mail:wangyp@yzu.edu.cn
ZHANG Wan-yu, XING Jun-qiang, SUN Qin-fu, WANG You-ping. Plant Regeneration and Genetic Transformation: From Empirical Exploration to Rational Design[J]. Biotechnology Bulletin, 2026, 42(10): 1-12.
递送系统 Delivery system | 主要原理 Principal mechanism | 核心优点 Key advantages | 主要局限/挑战 Major limitations/Challenges | 典型适用场景 Typical applications |
|---|---|---|---|---|
| 农杆菌介导 | 利用细菌天然机制将T-DNA整合至植物基因组 | 整合拷贝数低; 可转移大片段; 技术成熟、操作简便、成本较低 | 存在宿主偏好性,对部分单子叶及难转化物种效率低 | 多数双子叶及部分单子叶植物的稳定转化 |
| 基因枪轰击 | 通过高压气体将包裹DNA的金属微粒(金/钨)直接轰击入植物细胞或细胞器 | 几乎不受物种、基因型限制; 可递送多种形态物质; 能高效转化叶绿体和线粒体 | 整合拷贝数多且随机,易导致基因沉默或表达不稳定; 设备成本较高; 条件优化复杂; 可能造成组织物理损伤 | 小麦、水稻等单子叶作物的遗传转化; 细胞器转化 |
| 纳米载体递送 | 利用纳米材料(如碳纳米管、脂质体)包裹核酸,通过物理或化学作用穿透细胞壁和细胞膜递送基因 | 潜力上可适用于任何物种; 可递送核蛋白复合体,避免DNA整合; 避免核酸胞内降解,转化效率高 | 技术不成熟,缺乏标准化流程; 纳米材料的环境影响尚不明确 | CRISPR/Cas9等基因编辑工具的递送; 难转化物种(如林木、藻类)的遗传工程 |
| 病毒载体递送 | 利用改造后的植物病毒在体内移动和复制 | 快速产生大量目标蛋白,无需基因组整合; 不依赖组织培养 | 承载容量小; 通常不整合入基因组,外源基因无法稳定遗传; 存在生物安全风险 | 基因功能快速验证; 植物生物反应器生产药用蛋白; 基因编辑 |
Table 1 Comparison of major delivery systems for plant genetic transformation
递送系统 Delivery system | 主要原理 Principal mechanism | 核心优点 Key advantages | 主要局限/挑战 Major limitations/Challenges | 典型适用场景 Typical applications |
|---|---|---|---|---|
| 农杆菌介导 | 利用细菌天然机制将T-DNA整合至植物基因组 | 整合拷贝数低; 可转移大片段; 技术成熟、操作简便、成本较低 | 存在宿主偏好性,对部分单子叶及难转化物种效率低 | 多数双子叶及部分单子叶植物的稳定转化 |
| 基因枪轰击 | 通过高压气体将包裹DNA的金属微粒(金/钨)直接轰击入植物细胞或细胞器 | 几乎不受物种、基因型限制; 可递送多种形态物质; 能高效转化叶绿体和线粒体 | 整合拷贝数多且随机,易导致基因沉默或表达不稳定; 设备成本较高; 条件优化复杂; 可能造成组织物理损伤 | 小麦、水稻等单子叶作物的遗传转化; 细胞器转化 |
| 纳米载体递送 | 利用纳米材料(如碳纳米管、脂质体)包裹核酸,通过物理或化学作用穿透细胞壁和细胞膜递送基因 | 潜力上可适用于任何物种; 可递送核蛋白复合体,避免DNA整合; 避免核酸胞内降解,转化效率高 | 技术不成熟,缺乏标准化流程; 纳米材料的环境影响尚不明确 | CRISPR/Cas9等基因编辑工具的递送; 难转化物种(如林木、藻类)的遗传工程 |
| 病毒载体递送 | 利用改造后的植物病毒在体内移动和复制 | 快速产生大量目标蛋白,无需基因组整合; 不依赖组织培养 | 承载容量小; 通常不整合入基因组,外源基因无法稳定遗传; 存在生物安全风险 | 基因功能快速验证; 植物生物反应器生产药用蛋白; 基因编辑 |
嵌合体 Chimera | 转化物种 Transformed species | 外植体类型 Explant type | 效果 Performance | 参考文献 Reference |
|---|---|---|---|---|
| TaGRF4-GIF1 | 小麦、水稻、柑橘 | 幼胚、愈伤组织 | 显著提高再生效率,缩短再生时间,提高可转化基因型数量 | [ |
| TaGRF4-GIF1 | 小麦、大麦 | 幼胚 | 促进再生,但大麦中抑制再生,需优化表达 | [ |
TaGRF4-GIF1 ZmGRF1-GIF1 | 玉米 | 幼胚 | 与三元载体系统结合,小麦来源与玉米来源的GRF-GIF均大幅提高再生转化频率 | [ |
| TaGRF4-OsGIF1 | 高粱 | 幼胚 | 与三元载体系统结合,转化效率达38.28%,为原始方法的7.71倍,转化流程缩短 | [ |
TaGRF4-GIF1 DcGRF4-GIF1 | 铁皮石斛 | 叶片、茎段 | 显著提高植株再生效率 | [ |
| GRF5 | 甜菜、油菜、大豆、向日葵、玉米 | 愈伤组织、幼胚 | 提高再生与转化效率,克服基因型依赖性 | [ |
| ClGRF4-GIF1 | 西瓜 | 子叶、下胚轴 | 转化效率最高达67.27%,实现基因型非依赖性转化 | [ |
| SlGRF4c-GIF1a | 番茄 | 愈伤组织 | 显著提高再生效率,缩短再生周期约1个月 | [ |
TaGRF4-TaGIF1 VvGRF4-VvGIF1 CcGRF4-GIF1 | 二倍体野生草莓 | 叶片 | 三种来源的嵌合体均显著提高再生效率 | [ |
| AtGRF5 | 黄瓜、甜瓜、西瓜 | 子叶、愈伤组织 | 促进愈伤增殖与干细胞扩增,提高转化效率 | [ |
| GmGRF3-GIF1 | 大豆 | 子叶节 | 显著提高再生与转化效率,扩大可转化品种范围 | [ |
| AcGRF4-AcGIF1 | 猕猴桃 | 叶片、下胚轴、顶芽;叶柄 | 显著提高发根农杆菌介导的毛状根诱导率和再生效率 | [ |
BoGRF4-GIF-BoGRF4 BoGRF5-GIF-BoGRF5 | 甘蓝 | 子叶、下胚轴 | 均显著提高再生效率,BoGRF5-GIF-BoGRF5效果更强,再生效率提高55.2% | [ |
| GRF4-GIF1 | 莴苣 | 子叶 | 显著提升多个栽培及野生种的体外再生效率和不定芽发生频率 | [ |
| CpGRF4-CpGIF1 | 柑橘 | 上胚轴 | 植物再生率达44.17%,为空载对照的3倍 | [ |
Table 2 Application performance of GRF-GIF chimeras in plant regeneration and genetic transformation
嵌合体 Chimera | 转化物种 Transformed species | 外植体类型 Explant type | 效果 Performance | 参考文献 Reference |
|---|---|---|---|---|
| TaGRF4-GIF1 | 小麦、水稻、柑橘 | 幼胚、愈伤组织 | 显著提高再生效率,缩短再生时间,提高可转化基因型数量 | [ |
| TaGRF4-GIF1 | 小麦、大麦 | 幼胚 | 促进再生,但大麦中抑制再生,需优化表达 | [ |
TaGRF4-GIF1 ZmGRF1-GIF1 | 玉米 | 幼胚 | 与三元载体系统结合,小麦来源与玉米来源的GRF-GIF均大幅提高再生转化频率 | [ |
| TaGRF4-OsGIF1 | 高粱 | 幼胚 | 与三元载体系统结合,转化效率达38.28%,为原始方法的7.71倍,转化流程缩短 | [ |
TaGRF4-GIF1 DcGRF4-GIF1 | 铁皮石斛 | 叶片、茎段 | 显著提高植株再生效率 | [ |
| GRF5 | 甜菜、油菜、大豆、向日葵、玉米 | 愈伤组织、幼胚 | 提高再生与转化效率,克服基因型依赖性 | [ |
| ClGRF4-GIF1 | 西瓜 | 子叶、下胚轴 | 转化效率最高达67.27%,实现基因型非依赖性转化 | [ |
| SlGRF4c-GIF1a | 番茄 | 愈伤组织 | 显著提高再生效率,缩短再生周期约1个月 | [ |
TaGRF4-TaGIF1 VvGRF4-VvGIF1 CcGRF4-GIF1 | 二倍体野生草莓 | 叶片 | 三种来源的嵌合体均显著提高再生效率 | [ |
| AtGRF5 | 黄瓜、甜瓜、西瓜 | 子叶、愈伤组织 | 促进愈伤增殖与干细胞扩增,提高转化效率 | [ |
| GmGRF3-GIF1 | 大豆 | 子叶节 | 显著提高再生与转化效率,扩大可转化品种范围 | [ |
| AcGRF4-AcGIF1 | 猕猴桃 | 叶片、下胚轴、顶芽;叶柄 | 显著提高发根农杆菌介导的毛状根诱导率和再生效率 | [ |
BoGRF4-GIF-BoGRF4 BoGRF5-GIF-BoGRF5 | 甘蓝 | 子叶、下胚轴 | 均显著提高再生效率,BoGRF5-GIF-BoGRF5效果更强,再生效率提高55.2% | [ |
| GRF4-GIF1 | 莴苣 | 子叶 | 显著提升多个栽培及野生种的体外再生效率和不定芽发生频率 | [ |
| CpGRF4-CpGIF1 | 柑橘 | 上胚轴 | 植物再生率达44.17%,为空载对照的3倍 | [ |
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