Establishment and Application of the Genetic Transformation System for Hybrid Pennisetum‘Bangde No.1’

doi:10.11733/j.issn.1007-0435.2026.10.030

Acta Agrestia Sinica ›› 2026, Vol. 34 ›› Issue (10): 3853-3861.DOI: 10.11733/j.issn.1007-0435.2026.10.030

Previous Articles Next Articles

Establishment and Application of the Genetic Transformation System for Hybrid Pennisetum‘Bangde No.1’

LI Tian¹, WANG Ting-ting², LI Wei¹, GUAN Cong², WANG Hui¹

1. College of Grassland Science and Technology, China Agricultural University, Beijing 100193, China;
2. Institute of Leisure Agriculture, Shandong Academy of Agricultural Sciences, Jinan, Shandong Province 250100, China

Received:2026-06-04 Revised:2026-07-09 Online:2026-10-15 Published:2026-10-10

杂交狼尾草‘邦得1号’遗传转化体系的建立及应用

李甜¹, 王婷婷², 李薇¹, 管聪², 王慧¹

1. 中国农业大学/草业科学与技术学院, 北京 100193;
2. 山东省农业科学院/休闲农业研究所, 山东济南 250100

通讯作者: 王慧, E-mail: huiwang211@cau.edu.cn
作者简介:李甜（1998-），女，汉族，陕西宝鸡人，硕士研究生，主要从事草类植物分子育种研究，E-mail：ltbwcx9@163.com

Abstract

Abstract: Hybrid Pennisetum （Pennisetum glaucum×Pennisetum purpureum） is an important perennial forage crop in China’s livestock production with advantages such as high biomass yield， rapid growth， and strong adaptability. However， the absence of a stable and efficient genetic transformation system severely restricts the functional verification of stress-resistant genes and molecular breeding. In this study， hybrid Pennisetum ‘Bangde No. 1’ was used as material. Through the systematic screening of explants and surface disinfection methods， callus induction， and the optimization of key genetic transformation parameters including Agrobacterium strain， bacterial concentration， acetosyringone （AS） dosage， and infection methods， an Agrobacterium-mediated genetic transformation system was established. The results showed that surface sterilization with 75% ethanol combined with 7.5% sodium hypochlorite was optimal for seed disinfection， The optimal transformation conditions consisted of Agrobacterium tumefaciens strain EHA105， a bacterial suspension at OD₆₀₀=0.8， 150 μmol·L^-1 AS， glutamine pretreatment， and vacuum-assisted infection， resulting in a transformation efficiency of 33%. Using this system， positive transformants carrying the cold-tolerance genes ZmRR1 and PpICE1-1 were successfully obtained. These results indicate that optimizing infection conditions and improving embryogenic callus quality and resistant callus differentiation are critical for enhancing transformation efficiency. The established system provides technical support for functional studies of stress-resistance genes and the breeding of cold-tolerant hybrid Pennisetum cultivars.

Key words: Hybrid Pennisetum, Embryogenic callus tissue, Genetic transformation, Agrobacterium tumefaciens

摘要： 杂交狼尾草（Pennisetum glaucum×Pennisetum purpureum）具有生物量大、生长快、适应性强等优点，是我国畜牧生产中重要的多年生牧草。然而，其稳定高效的遗传转化体系尚未建立，严重制约了其抗逆基因功能解析与定向分子育种进程。为建立稳定高效的杂交狼尾草遗传转化体系，本研究以 ‘邦得1号’为材料，系统筛选外植体及其消毒方式、愈伤组织诱导条件、农杆菌菌株、菌液浓度、乙酰丁香酮（Acetosyringone， AS）浓度及侵染方式。结果表明，种子经75%乙醇和7.5%次氯酸钠处理后可获得无菌外植体并诱导胚性愈伤组织；采用EHA105、菌液OD₆₀₀为0.8、AS浓度150 μmol·L^-1，并结合谷氨酰胺（Glutamine， Gln）预处理和负压侵染，转化效率达33%。利用该体系成功获得导入ZmRR1、PpICE1-1等耐寒基因的阳性转化株。综上，优化侵染条件、提高胚性愈伤组织质量和抗性愈伤分化率，是提升转化效率的关键。该体系为杂交狼尾草抗逆基因功能研究和抗寒新品种选育提供技术支撑。

关键词: 杂交狼尾草, 胚性愈伤组织, 遗传转化, 根癌农杆菌

CLC Number:

LI Tian, WANG Ting-ting, LI Wei, GUAN Cong, WANG Hui. Establishment and Application of the Genetic Transformation System for Hybrid Pennisetum‘Bangde No.1’[J]. Acta Agrestia Sinica, 2026, 34(10): 3853-3861.

李甜, 王婷婷, 李薇, 管聪, 王慧. 杂交狼尾草‘邦得1号’遗传转化体系的建立及应用[J]. 草地学报, 2026, 34(10): 3853-3861.

References

[1] CHEN Z T，HE S L，HUANG Y B. Research advances in forage Pennisetum resources[J]. Acta Agrestia Sinica，2010，18（5）：740-748 陈志彤，何水林，黄毅斌. 狼尾草属牧草研究进展[J]. 草地学报，2010，18（5）：740-748
[2] HOU X C，TENG K，GUO Q，et al. Research advances in forage Pennisetum resource[J]. Bulletin of Botany，2022，57（6）：814-825 侯新村，滕珂，郭强，等. 狼尾草属牧草研究进展[J]. 植物学报，2022，57（6）：814-825
[3] WANG W Q，ZHOU H L，TANG J. Research advances on development and utilization of forage of Pennisetum spp[J]. Chinese Journal of Tropical Agriculture，2018，38（6）：49-55，78 王文强，周汉林，唐军. 狼尾草属牧草研究及利用进展[J]. 热带农业科学，2018，38（6）：49-55，78
[4] RASHID M，AMJAD N，SHAH K H. Chlorophyll a fluorescence kinetics uncover differential photosystem II response and salinity tolerance mechanisms in pearl millet （Pennisetum glaucum L.）[J]. BMC Plant Biology，2026，26（1）：277
[5] ABAGISA R A，ABEBE Y A，OBSA B T，et al. Assessing agronomic practices and constraints of pearl millet [Pennisetum glaucum （L.） R. Br.] production in semiarid areas of the west hararghe zone，eastern Ethiopia[J]. International Journal of Agronomy，2026，2026（1）：8437238
[6] YAN H D，SUN M，ZHANG Z R，et al. Pangenomic analysis identifies structural variation associated with heat tolerance in pearl millet[J]. Nature Genetics，2023，55（3）：507-518
[7] LIN T，LI B Y，LIU L S，et al. Silage technology of hybrid Pennisetum in southwest China[J]. China Animal Industry，2025（1）：54-55 林涛，李冰洋，刘卢生，等. 西南地区杂交狼尾草青贮技术[J]. 中国畜牧业，2025（1）：54-55
[8] LIN S X，GAO C F，ZHANG X P，et al. Research progress on comprehensive utilization of hybrid Pennisetum[J]. Fujian Journal of Animal Husbandry and Veterinary Medicine，2011，33（1）：32-33 林仕欣，高承芳，张晓佩，等. 杂交狼尾草综合利用的研究进展[J]. 福建畜牧兽医，2011，33（1）：32-33
[9] WANG P Q，DUAN C R，WANG B C，et al. Study of Pennisetum pureum on function of soil and water conservation[J]. Journal of Soil and Water Conservation，2005，19（1）：114-116，135 王凭青，段传人，王伯初，等. 杂交狼尾草水土保持能力的实验研究[J]. 水土保持学报，2005，19（1）：114-116，135
[10] CHEN J，ZHUO G H，WANG S，et al. Study on remediation mechanism of Pennisetum in typical cadmium contaminated soil in South China[J]. Straits Science，2019（4）：29-31，38 陈锦，卓桂华，王珊，等. 狼尾草属植物在南方典型镉污染土壤修复机制研究[J]. 海峡科学，2019（4）：29-31，38
[11] XUE C Y，HE W M，ZHANG R L. The feasibility study of pulping and papermaking with hybridized Chinese Pennisetum[J]. China Pulp & Paper Industry，2006，27（1）：69-72 薛崇昀，贺文明，张睿玲. 杂交狼尾草制浆造纸的可行性研究[J]. 中华纸业，2006，27（1）：69-72
[12] ZHANG J Z. Intercropping hybrid Pennisetum in mountain orchards and its benefit analysis[J]. Fujian Fruits，2005（1）：58-59 张菊珠. 山地果园套种杂交狼尾草技术及其效益分析[J]. 福建果树，2005（1）：58-59
[13] CHEN Z T，YING C Y，LIN Y S，et al. Cultivation techniques and utilization value of hybrid Pennisetum[J]. Fujian Agricultural Science and Technology，2006，37（2）：44-45 陈志彤，应朝阳，林永生，等. 杂交狼尾草的栽培技术与利用价值[J]. 福建农业科技，2006，37（2）：44-45
[14] HIEI Y，OHTA S，KOMARI T，et al. Efficient transformation of rice （Oryza sativa L.） mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA[J]. The Plant Journal，1994，6（2）：271-282
[15] CAO X S，XIE H T，WANG Z H，et al. An efficient tissue-culture-free soybean genetic transformation technology using the extremely simple cut-dip-budding strategy[J]. The Innovation，2026，7（3）：101221
[16] WANG P Q，DUAN C R，WANG B C，et al. Tissue culture with different organs of Pennisetum pureum[J]. Journal of Chongqing University，2005（6）：118-120 王凭青，段传人，王伯初，等. 杂交狼尾草不同外植体材料组织培养实验[J]. 重庆大学学报（自然科学版），2005（6）：118-120
[17] GONG S F，ZHANG H L，ZHANG X Y，et al. Asepsis sowing and tissure culture of Pennisetum alopecuroides （L.） spreng[J]. Plant Physiology Communications，2010，46（6）：611-612 龚束芳，张瀚俪，张晓莹，等. 狼尾草成熟种子的无菌播种与组织培养[J]. 植物生理学通讯，2010，46（6）：611-612
[18] ZANG W J，CHEN Y，LI Q，et al. Tissue culture differentiation and callus induction of different explant from Pennisetum americanum×P.purureum[J]. Pratacultural Science，2015，32（9）：1451-1456 臧文静，陈莹，李青，等. 杂交狼尾草不同外植体愈伤组织诱导[J]. 草业科学，2015，32（9）：1451-1456
[19] MU T，ZHANG X Y，WANG J G，et al. The optimization of Pennisetum agrobacterium-mediated transformation system and obtaining of transgenic plants[J]. Crops，2013（1）：45-48，158 牟彤，张晓莹，王金刚，等. 狼尾草农杆菌转化体系的优化和转基因植株的获得[J]. 作物杂志，2013（1）：45-48，158
[20] ACHARY V M M，HERNANDEZ L R B，GAO H R，et al. High-throughput genetic transformation and genome editing in pearl millet （Pennisetum glaucum L.）[J]. Plant Physiology，2025，199（3）：kiaf499
[21] GIRGI M，O’KENNEDY M M，MORGENSTERN A，et al. Transgenic and herbicide resistant pearl millet （Pennisetum glaucum L.） R.Br. via microprojectile bombardment of scutellar tissue[J]. Molecular Breeding，2002，10（4）：243-252
[22] O’KENNEDY M M，BURGER J T，BOTHA F C. Pearl millet transformation system using the positive selectable marker gene phosphomannose isomerase[J]. Plant Cell Reports，2004，22（9）：684-690
[23] GOLDMAN J J，HANNA W W，FLEMING G，et al. Fertile transgenic pearl millet [Pennisetum glaucum （L.） R. Br.] plants recovered through microprojectile bombardment and phosphinothricin selection of apical meristem-，inflorescence-，and immature embryo-derived embryogenic tissues[J]. Plant Cell Reports，2003，21（10）：999-1009
[24] GUO X M，CHE X M，PEI Y H，et al. Effect of acetosyringone on genetic transformation of receptor system of different waxy maizes[J]. Southwest China Journal of Agricultural Sciences，2013，26（3）：899-902 郭新梅，车昕明，裴玉贺，等. 乙酰丁香酮对不同糯玉米受体系统遗传转化的影响[J]. 西南农业学报，2013，26（3）：899-902
[25] WANG Z J，LI M X，ZHANG W J. Optimization of Agrobacterium tumefaciens-mediated genetic transformation system of alfalfa[J]. Acta Agrestia Sinica，2024，32（6）：1665-1671 王之杰，李明序，张万军. 根癌农杆菌介导的紫花苜蓿遗传转化体系的优化[J]. 草地学报，2024，32（6）：1665-1671
[26] CHENG M，FRY J E，PANG S，et al. Genetic transformation of wheat mediated by Agrobacterium tumefaciens[J]. Plant Physiology，1997，115（3）：971-980
[27] LIU Y R，CEN H F，YAN J P，et al. Optimizing of Agrobacterium-mediated transformation of switchgrass cultivars[J]. Scientia Agricultura Sinica，2016，49（1）：80-89 刘燕蓉，岑慧芳，严建萍，等. 农杆菌介导的柳枝稷遗传转化体系的优化[J]. 中国农业科学，2016，49（1）：80-89
[28] TANG F，XU B，SHI F L，et al. Establishment of efficient tissue culture and regeneration system of Agropyron mongolicum Keng root tip[J]. Chinese Journal of Grassland，2021，43（3）：113-118 唐芳，徐舶，石凤翎，等. 蒙古冰草根尖高效组培再生体系的建立[J]. 中国草地学报，2021，43（3）：113-118
[29] FAN Y H，XU Y，ZHU S C，et al. Construction of tissue culture regeneration system of Pennisetum glaucum[J]. Acta Agrestia Sinica，2023，31（1）：89-95 范欲航，许悦，祝诗词，等. 美洲狼尾草组织培养再生体系的建立[J]. 草地学报，2023，31（1）：89-95
[30] LI R Y，QU R D. High throughput Agrobacterium-mediated switchgrass transformation[J]. Biomass and Bioenergy，2011，35（3）：1046-1054
[31] HUANG B Y，ZHUO R Y，QIAO G R. Comparative analysis of different types of calli in moso bamboo[J]. Journal of Nanjing Forestry University （Natural Sciences Edition），2023，47（6）：141-149 黄碧芸，卓仁英，乔桂荣. 毛竹不同类型愈伤组织比较分析[J]. 南京林业大学学报， 2023， 47（6）： 141-149
[32] WANG Q Y，CHENG H Y，WANG W Q，et al. Efficient induction of callus and plant regeneration from Miscanthus sacchariflorus young ear[J]. Plant Diversity and Resources，2013，35（2）：171-172 王青云，程红焱，王伟青，等. 芒属植物荻愈伤组织的诱导与植株再生[J]. 植物分类与资源学报，2013，35（2）：171-172
[33] OZAWA K，KAWAHIGASHI H，KAYANO T，et al. Enhancement of regeneration of rice （Oryza sativa L.） calli by integration of the gene involved in regeneration ability of the callus[J]. Plant Science，2003，165（2）：395-402
[34] SAHOO K K，TRIPATHI A K，PAREEK A，et al. An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars[J]. Plant Methods，2011，7（1）：49
[35] MIN Y，JI Q L，WANG X H，et al. Regeneration of Populus euphratica via organogenesis and embryogenesis[J]. Biotechnology，2007，17（6）：52-55 闵义，计巧灵，王雪华，等. 胡杨器官和体胚发生方式的植株再生[J]. 生物技术，2007，17（6）：52-55
[36] YI Z L，CAO S Y，WANG L，et al. Improvement of transformation frequency of rice mediated by Agrobacterium[J]. Acta Genetica Sinica，2001，28（4）：352-358
[37] YU G R，LIU Y，DU W P，et al. Optimization of Agrobacterium tumefaciens-mediated immature embryo transformation system and transformation of glyphosate-resistant gene 2mG2-EPSPS in maize （Zea mays L.）[J]. Journal of Integrative Agriculture，2013，12（12）：2134-2142
[38] SONG G Q，WALWORTH A，HANCOCK J F. Factors influencing Agrobacterium-mediated transformation of switchgrass cultivars[J]. Plant Cell，Tissue and Organ Culture （PCTOC），2012，108（3）：445-453
[39] KHAN S，FAHIM N，SINGH P，et al. Agrobacterium tumefaciens mediated genetic transformation of Ocimum gratissimum： A medicinally important crop[J]. Industrial Crops and Products，2015，71：138-146
[40] TRICK H N，FINER J J. Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max （L.） Merrill] embryogenic suspension culture tissue[J]. Plant Cell Reports，1998，17（6）：482-488
[41] TRICK H N，FINER J J. SAAT： sonication-assisted Agrobacterium-mediated transformation[J]. Transgenic Research，1997，6（5）：329-336
[42] BIDNEY D，SCELONGE C，MARTICH J，et al. Microprojectile bombardment of plant tissues increases transformation frequency by Agrobacterium tumefaciens[J]. Plant Molecular Biology，1992，18（2）：301-313
[43] ZHANG W J，DEWEY R E，BOSS W，et al. Enhanced Agrobacterium-mediated transformation efficiencies in monocot cells is associated with attenuated defense responses[J]. Plant Molecular Biology，2013，81（3）：273-286
[44] LOWE K，WU E，WANG N，et al. Morphogenic regulators baby boom and wuschel improve monocot transformation[J]. The Plant Cell，2016，28（9）：1998-2015
[45] DEBERNARDI J M，TRICOLI D M，ERCOLI M F，et al. A GRF-GIF chimeric protein improves the regeneration efficiency of transgenic plants[J]. Nature Biotechnology，2020，38（11）：1274-1279

Establishment and Application of the Genetic Transformation System for Hybrid Pennisetum‘Bangde No.1’

杂交狼尾草‘邦得1号’遗传转化体系的建立及应用

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Yu-juan, HOU Bo, LIU Jun-qi, ZHANG Yun-wei, FAN Yan, BI Xiao-jing. Mutagenic Effects and Screening of Cold-Tolerant Germplasm of Hybrid Pennisetum under ARTP-EMS Combined Mutagenesis [J]. Acta Agrestia Sinica, 2026, 34(10): 3653-3661.
[2]	LI Xin-ru, SU Kun-long, ZHANG Xin-yu, MA Li-chao, LI Shuo, YU Qian-qian, JIANG Teng-hui, WANG Yu-rou, YANG Guo-feng. Establishment of High-Efficiency Tissue Regeneration and Genetic Transformation System for Leymus chinensis [J]. Acta Agrestia Sinica, 2026, 34(10): 3862-3874.
[3]	FAN Bo-bo, LIU Jie, WU Jing, YAN Xiu-xiu, MA Yan-hong. Establishment of A Seed Induction Regeneration System for Agropyron mongolicum Keng and Study of Agrobacterium-mediated Transformation [J]. Acta Agrestia Sinica, 2026, 34(10): 3875-3881.
[4]	HUANG Hai-jun, ZHANG Yu-qi, DU Wen-xuan, KONG Wei-ye, SHEN Guo-an, LIU Ya-ling, PANG Yong-zhen. Research Progress on Germplasm Resources of Sainfoin (Onobrychis viciaefolia) [J]. Acta Agrestia Sinica, 2024, 32(8): 2346-2356.
[5]	WANG Zhi-jie, LI Ming-xu, ZHANG Wan-jun. Optimization of Agrobacterium Tumefaciens-Mediated Genetic Transformation System of Alfalfa [J]. Acta Agrestia Sinica, 2024, 32(6): 1665-1671.
[6]	LI Qi-dong, LI Cui, CHEN Zhi-min, ZHANG Li-xiang, GUO Qiang. Exploration on the Functions of IlNRAMP5 Mediating the Cd²⁺ uptake in Iris lactea [J]. Acta Agrestia Sinica, 2023, 31(6): 1640-1647.
[7]	TIAN Shu-ting, CAO Xiao-yun, SHI Chun-ying, DU Ling-juan. Optimization of Agrobacterium Tumefaciens-Mediated Genetic Transformation System based on Callus of Grape Hyacinth [J]. Acta Agrestia Sinica, 2023, 31(4): 1234-1241.
[8]	ZHAO Xing-kun, SUN Hao, YANG Li-yun, LUO Li-juan, LIU Pan-dao. Establishment of the Genetic Transformation System of Stylosanthes gracilis mediated with Agrobacterium rhizogenes [J]. Acta Agrestia Sinica, 2023, 31(2): 581-586.
[9]	CAO Xiao-yu, WANG Xue-min, GUO Ji-cheng, CUI Miao-miao, ZHANG Jin-jin, LIU Jiao-jiao, HE Jun-ying. Cloning,Expression Characteristics and Genetic Transformation Analysis of A bZIP Transcription Factor MsbZIP1 in Alfalfa [J]. Acta Agrestia Sinica, 2020, 28(3): 613-622.
[10]	MU Zhi-xin, SHI Ying, ZHANG Li-jun, ZHOU Jian-ping. Introduction Of OsVTE Gene Enhanced Osmotic Stress Tolerance of Transgenic Chicory Plants [J]. Acta Agrestia Sinica, 2017, 25(4): 839-844.
[11]	CHEN Xin-zhu, GAO Cheng-fang, ZHANG Xiao-pei, LI Wen-yang, WENG Bo-qi. Effects of Molasses on the Fermentation Quality of Mixed Silage of Ramie and Hybrid Pennisetum [J]. Acta Agrestia Sinica, 2016, 24(6): 1358-1362.
[12]	LIU Da-lin, SUN Qi-xin, SHAO Jiang, CHEN Ming-hui. Effects of Different Soil Cd Concentrations on Physiological Characteristics of Two Pennisetums at Different Growth Stages [J]. Acta Agrestia Sinica, 2016, 24(4): 854-858.
[13]	LI Zhen-yi, LONG Rui-cai, ZHANG Tie-jun, YANG Qing-chuan, KANG Jun-mei. Cloning and Functional Analysis of MsHSP 17.7 Gene from Alfalfa [J]. Acta Agrestia Sinica, 2016, 24(1): 121-128.
[14]	ZHANG Li, GAN Xiao-yan, SHI Lei, CHEN Yu-chao, NIE Feng-jie, SONG Yu-xia. Transformation System of Medicago sativa ‘Xinjiang Daye’ with Betaine Aldehyde Dehydrogenase Gene [J]. , 2014, 22(2): 359-365.
[15]	ZHANG Li-jun, CHENG Lin-mei, DU Jian-zhong, WANG Yi-xue, HAO Yao-shan, LI Gui-quan, SUN Yi. Cotton APX Gene Enhances Stress Tolerance of Cichorium intybus [J]. , 2012, 20(1): 152-158.