High Temperature and Trifluralin Induce 2n Pollen in Hydrangea macrophylla

doi:10.16420/j.issn.0513-353x.2024-0950

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (11): 3090-3102.doi: 10.16420/j.issn.0513-353x.2024-0950

• New Technology and New Method • Previous Articles Next Articles

High Temperature and Trifluralin Induce 2n Pollen in Hydrangea macrophylla

YANG Ziwei¹, ZHANG Hua¹, QIU Shuai², TANG Yuchao¹, FU Xuehao¹, ZHANG Gachen¹, SUN Ming¹^,^*(), and LIU Xiuli¹^,^*()

¹ State Key Laboratory of Efficient Production of Forest Resources， Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education，Engineering Research Center of Landscape Environment of Ministry of Education，Beijing Laboratory of Urban and Rural Ecological Environment，National Engineering Research Center for Floriculture，Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding，School of Landscape Architecture，Beijing Forestry University， Beijing 100083， China
² Hangzhou Landscaping Incorporated， Hangzhou 310020， China

Received:2025-01-08 Revised:2025-08-06 Online:2025-11-26 Published:2025-11-26
Contact: SUN Ming, and LIU Xiuli

Abstract

Abstract:

This study observed the meiotic process of microsporoblasts using flat-topped Hydrangea macrophylla‘Ziye’as material，and then treated flower buds with trifluralin wrapping and high temperature treatments at different meiotic periods. This study subsequently determined the 2n pollen induction rate of the different treatments and the viability of the 2n pollen in vitro germination. The results demonstrated that the trifluralin wrapping treatment achieved the highest induction effect during the“pollen mother cell”stage，while high-temperature induction exhibited optimal efficacy from the“late meiotic phase to metaphaseⅠ”. In the trifluralin induction process，a 48 h exposure to 0.1% trifluralin in conjunction with 2% dimethyl sulfoxide yielded optimal results. For high-temperature induction，treatment at 40 ℃ for 6 h was the ideal condition. The 2n pollen produced by both induction methods retained viability，making it suitable for subsequent hybridization experiments. In conclusion，both trifluralin and high temperature treatments can successfully induce the formation of 2n pollen，with trifluralin being more efficient.

Key words: Hydrangea macrophylla, 2n pollen induction, chromosome doubling, trifluralin, high temperature induction

YANG Ziwei, ZHANG Hua, QIU Shuai, TANG Yuchao, FU Xuehao, ZHANG Gachen, SUN Ming, and LIU Xiuli. High Temperature and Trifluralin Induce 2n Pollen in Hydrangea macrophylla[J]. Acta Horticulturae Sinica, 2025, 52(11): 3090-3102.

Figures/Tables 12

References 33

[1]	Adams K L, Wendel J F. 2005. Polyploidy and genome evolution in plants. Current Opinion in Plant Biology,8:135-141.
[2]	Alexander L. 2020. Ploidy level influences pollen tube growth and seed viability in interploidy crosses of Hydrangea macrophylla. Frontiers in Plant Science,11:100.
[3]	Bretagnolle F, Thompson J D. 1995. Gametes with the somatic chromosome number:mechanisms of their formation and role in the evolution of autopolyploid plants. New Phytologist,129:1-22.
[4]	Broughton S, Castello M, Liu L, Killen J, Hepworth A, O’Leary R. 2020. The effect of caffeine and trifluralin on chromosome doubling in wheat anther culture. Plants, 9 (1):105.
[5]	Cheng Shiping, Yao Pengqiang, Geng Xining, Liu Chunyang, Xie Lihua. 2022. High temperature treatment generates unreduced pollen in Paeonia suffruticosa. Acta Horticulturae Sinica, 49 (3):581-589. (in Chinese)
	程世平, 姚鹏强, 耿喜宁, 刘春洋, 谢丽华. 2022. 高温诱导牡丹产生未减数花粉. 园艺学报, 49 (3):581-589.
[6]	Chu Lihong, Peng Jiajia, Wang Zhao, Zhao Kai, Yuan Suxia, Ming Jun, Liu Chun. 2014. Studies on polyploid induction in Anthurium andraeanum using oryzalin,trifluralin and colchicine. Acta Horticulturae Sinica, 41 (11):2275-2280. (in Chinese)
	储丽红, 彭佳佳, 王钊, 赵凯, 袁素霞, 明军, 刘春. 2014. 氨磺灵、氟乐灵和秋水仙素诱导安祖花多倍体的研究. 园艺学报, 41 (11):2275-2280.
[7]	Crespel L, Gudin S, Meynet J, Zhang D. 2002. AFLP-based estimation of 2n gametophytic heterozygosity in two parthenogenetically derived dihaploids of Rosa hybrida L. Theoretical and Applied Genetics, 104 (2-3):451-456.
[8]	Dewitte A, Eeckhaut T, Van Huylenbroeck J, van Bockstaele E. 2010. Induction of 2n pollen formation in Begonia by trifluralin and N₂O treatments. Euphytica, 171 (2):283-293.
[9]	Gao Peng, Lin Wei, Kang Xiangyang. 2004. Pollen chromosome doubling of Eucommia ulmoides induced by colchicine. Journal of Beijing Forestry University,(4):39-42. (in Chinese)
	高鹏, 林威, 康向阳. 2004. 秋水仙碱诱导杜仲花粉染色体加倍的研究. 北京林业大学学报,(4):39-42.
[34]	Wang Aihua, Wu Qingqing, Yang Lan, Chen Zhilin. 2017. Study on 2n pollen induction and cytological mechanism of 2n pollen formation in Dendrobium officinale. Acta Botanica Boreali-Occidentalia Sinica, 37 (2):273-277. (in Chinese)
	王爱华, 吴青青, 杨澜, 陈之林. 2017. 铁皮石斛2n花粉诱导及其形成的细胞机制研究. 西北植物学报, 37 (2):273-277.
[35]	Wang Yulin, Huang Kun, Zhou Yiqing, Su Xue, Li Bin. 2024. Study on cold resistance of three Hydrangea species in Beijing during natural overwintering. Journal of Anhui Agricultural Sciences, 52 (17):42-45. (in Chinese)
	王煜琳, 黄焜, 周祎晴, 苏雪, 李斌. 2024. 自然降温过程中北京地区3种绣球的耐寒性研究. 安徽农业科学, 52 (17):42-45.
[36]	Wang Ziyue, Liu Guangxin, Qiu Shuai, Gao Kai, Wei Jianfen, Zhen Yan, Xi Mengli. 2023. Chromosome behaviors during the meiosis of microspore mother cells and pollen characteristics in Hydrangea. Scientia Silvae Sinicae, 59 (7):89-95. (in Chinese)
	王子玥, 刘光欣, 邱帅, 高凯, 魏建芬, 甄艳, 席梦利. 2023. 绣球小孢子母细胞减数分裂染色体行为及花粉特征. 林业科学, 59 (7):89-95.
[37]	Wei Zhaofen. 1994. Revision of the genus Hydrangea in China. Guihaia,(2):101-121. (in Chinese)
	卫兆芬. 1994. 中国绣球属植物的修订. 广西植物,(2):101-121.
[38]	Wu Kepeng, Xu Weijie, Xie Lili, Pan Chongren. 2020. Introduction and adaptability of Hydrangea L. plants. Journal of Zhejiang Agricultural Sciences, 61 (3):549-551. (in Chinese)
	吴可鹏, 徐炜杰, 谢丽丽, 潘崇仁. 2020. 绣球属植物引种及适应性试验. 浙江农业科学, 61 (3):549-551.
[39]	Xin Haoyang, Wang Shuai, Liu Guangxin, Zhen Yan, Shi Jisen, Kuang Hualin, Xi Mengli. 2016. Relationship between the meiosis processes of microsporocytes and morphology of male flower buds and anthers in Populus deltoides‘Marsh’. Journal of Nanjing Forestry University(Natural Sciences Edition), 40 (2):48-52. (in Chinese)
	辛昊阳, 王帅, 刘光欣, 甄艳, 施季森, 匡华琳, 席梦利. 2016. 美洲黑杨小孢子母细胞减数分裂进程与花芽及花药外观形态相关性研究. 南京林业大学学报(自然科学版), 40 (2):48-52.
[40]	Zhang Hailiang, Song Jie, Li Yefang, Meng Jing, Guan Wenling. 2024. Studies on floral traits and meiosis process of pollen mother cells in Bougainvillea glabra‘Elizabeth Angus’. Molecular Plant Breeding,1-19. (in Chinese)
	张海立, 宋杰, 李叶芳, 孟静, 关文灵. 2024. 三角梅‘安格斯’花部特征及花粉母细胞减数分裂进程研究. 分子植物育种,1-19.
[41]	Zhang Pingdong. 2023. Research progress on the mechanism of 2n gamete occurrence in plants. Journal of Beijing Forestry University, 45 (7):1-8. (in Chinese)
	张平冬. 2023. 植物2n配子发生机理研究进展. 北京林业大学学报, 45 (7):1-8.
[42]	Zhao Chenguang, Li Yanjie, Zhang Pingdong, Kang Xiangyang. 2015. Cytological mechanism of unreduced 2n pollen formation in Populus euphratica. Journal of Nanjing Forestry University(Natural Sciences Edition), 39 (6):40-44. (in Chinese)
[10]	Gao S, Sun Y, Zhou Y. 2021. Colchicine induction of‘Old Blush’2n pollen for the hybridization and breeding of tetraploid rose. PeerJ,9:doi:10.7717/peerj.11043.
[11]	Hansen A L, Gertz A, Joersbo M, Andersen S B. 1998. Anti-microtubule herbicides for in vitro chromosome doubling in Beta vulgaris L. ovule culture. Euphytica, 101 (2):231-237.
[12]	Harder L D, Thomson J D. 1989. Evolutionary options for maximizing pollen dispersal of animal pollinated plants. The American Naturalist, 133 (3):323-344.
[13]	Heckmann S, Schubert V, Houben A. 2014. Holocentric plant meiosis:first sisters,then homologues. Cell Cycle, 13 (23):3623-3624.
[14]	Hempel P, Hohe A, Tränkner C. 2018. Molecular reconstruction of an old pedigree of diploid and triploid Hydrangea macrophylla genotypes. Frontiers in Plant Science,9:429.
[15]	Jiao Jun, Liang Lijun, Ma Hui, Li Fengtong, Chen Xiulan, He Rong. 2016. Introduction and cultivation technology of Hydrangea germplasm resources. Modern Agricultural Science and Technology,11:203-204. (in Chinese)
	焦隽, 梁丽君, 马辉, 李风童, 陈秀兰, 何榕. 2016. 绣球种质资源引进与栽培技术. 现代农业科技,11:203-204.
[16]	Jones K D, Reed S M, Rinehart T A. 2007. Analysis of ploidy level and its effects on guard cell length,pollen diameter,and fertility in Hydrangea macrophylla. HortScience, 42 (3):483-488.
[17]	Kang Xiangyang. 2010. Some understandings on polyploid breeding of poplars. Journal of Beijing Forestry University, 32 (5):149-153. (in Chinese)
	康向阳. 2010. 关于杨树多倍体育种的几点认识. 北京林业大学学报, 32 (5):149-153.
[18]	Kreiner J M, Kron P, Husband B C. 2017. Evolutionary dynamics of unreduced gametes. Trends in Genetics, 33 (9):583-593.
[19]	Li Yue, Kang Xiangyang, Yang Weiguang, Guo Changhua. 2008. Pollen chromosome doubling of Orychophragmus violaceus L. induced by high temperature. Acta Botanica Boreali-Occidentalia Sinica,(6):1128-1133. (in Chinese)
[42]	赵晨光, 李燕杰, 张平冬, 康向阳. 2015. 胡杨未减数2n花粉形成的细胞学机制. 南京林业大学学报(自然科学版), 39 (6):40-44.
[43]	Zhao Yifan, Kong Bo, Cheng Xutong, Li Liang, Ling Aoyu, Li Zhiqun, Kang Xiangyang, Zhang Pingdong. 2023. Pollen chromosome doubling induced by gibberellin spraying treatment and its effect on microtubule cytoskeleton in Populus bolleana. Journal of Beijing Forestry University, 45 (1):40-50. (in Chinese)
	赵一帆, 孔博, 程雪桐, 李亮, 凌傲宇, 李智群, 康向阳, 张平冬. 2023. 赤霉素喷洒处理诱导新疆杨2n花粉产生及其对微管骨架的影响. 北京林业大学学报, 45 (1):40-50.
[44]	Zhou Q, Cheng X, Kong B, Zhao Y, Li Z, Sang Y, Wu J, Zhang P. 2022. Heat shock-induced failure of meiosis I to meiosis II transition leads to 2n pollen formation in a woody plant. Plant Physiology, 189 (4):2110-2127.
[45]	Zhou Qing, Wu Jian, Sang Yaru, Zhao Zhengyang, Liu Meiqin, Zhang Pingdong. 2020. Pollen chromosome doubling induced by colchicine treatment andcreation of hybrid triploids in Populus canescens. Journal of Beijing Forestry University, 42 (3):119-126. (in Chinese)
	周晴, 吴剑, 桑亚茹, 赵征洋, 刘美芹, 张平冬. 2020. 秋水仙碱处理诱导银灰杨2n花粉与杂种三倍体创制. 北京林业大学学报, 42 (3):119-126.
[46]	Zlesak D C, Thill C A, Anderson N O. 2005. Trifluralin-mediated polyploidization of Rosa chinensis minima(Sim)Voss seedlings. Euphytica, 141 (3):281-290.
[19]	李悦, 康向阳, 杨伟光, 郭长花. 2008. 高温诱导诸葛菜花粉染色体加倍研究. 西北植物学报,(6):1128-1133.
[20]	Liao Xiaoshan, Wu Qingqing, Zhang Chaojun, Zhou Jun, Zheng Sixiang, Wang Linyu. 2016. Study on 2n pollen Induction and Its identification of Lilium oriental. Northern Horticulture,( 8):56-60. (in Chinese)
	廖晓珊, 吴青青, 张朝君, 周俊, 郑思乡, 王淋玉. 2016. 东方百合2n花粉诱导及鉴定研究. 北方园艺,(8):56-60.
[21]	Lim K B, Shen T M, Barba-Gonzalez R, Ramanna M S, van Tuyl J M. 2004. Occurrence of SDR 2n-gametes in Lilium Hybrids. Breeding Science, 54 (1):13-18.
[22]	Liu Guoyu, Wang Qing, Zhang Wenbo, Liu Ancheng, Zhao Xueyan, Li Yan. 2024. In vitro germination study of pollen from four Hydrangea species. Acta Agriculturae Boreali-Occidentalis Sinica, 33 (9):1681-1689. (in Chinese)
	刘国宇, 王庆, 张文波, 刘安成, 赵雪艳, 李艳. 2024. 4种绣球花粉离体萌发研究. 西北农业学报, 33 (9):1681-1689.
[23]	Mai Yini, Li Shuzhan, Suo Yujing, Sun Peng, Han Weijuan, Diao Songfeng, Wang Liyuan, Li Huawei, Fu Jianmin. 2019. Identification of natural 2n pollens in different persimmon germplasms and ascertainment of their induction period. Journal of China Agricultural University, 24 (12):44-52. (in Chinese)
	买旖旎, 李树战, 索玉静, 孙鹏, 韩卫娟, 刁松锋, 王丽媛, 李华威, 傅建敏. 2019. 柿不同种质中的天然2n花粉得率的评价及诱导时期的判定. 中国农业大学学报, 24 (12):44-52.
[24]	Mao Yanke, Zhang Pingdong, Shi Le, Dong Chunbo, Suo Yujing, Kang Xiangyang. 2013. The optimum conditions for inducing 2n pollen chromosome doubling by high temperature in Eucommia ulmoides. Journal of Beijing Forestry University, 35 (1):53-58. (in Chinese)
	毛彦科, 张平冬, 石乐, 董春波, 索玉静, 康向阳. 2013. 高温诱导杜仲2n花粉最佳处理条件研究. 北京林业大学学报, 35 (1):53-58.
[25]	Orjeda G, Freyre R, Iwanaga M. 1990. Production of 2n pollen in diploid Ipomoea trifida,a putative wild ancestor of sweetpotato. Journal of Heredity, 81 (6):462-467.
[26]	Ramanna M S, Jacobsen E. 2003. Relevance of sexual polyploidization for crop improvement-a review. Euphytica,133:3-18.
[27]	Ramsey J, Schemske D W. 1998. Pathways,mechanisms,and rates of polyploid formation in flowering plants. Annual Review of Ecology & Systematics,29:467-501.
[28]	Sattler M C, Carvalho C R, Clarindo W R. 2015. The polyploidy and its key role in plant breeding. Planta, 243 (2):281-296.
[29]	Sun Rongze, Zhu Shaocai, Cui Yaqi, Zhao Jiageng, Wang Yuxuan, Yu Xiaonan. 2024. Megagametogenesis and 2n megagamete induction in Paeonia lactiflora. Journal of Northeast Forestry University, 52 (5):49-55. (in Chinese)
	孙榕泽, 朱绍才, 崔雅琦, 赵家庚, 王宇暄, 于晓南. 2024. 芍药雌配子发育和2n雌配子诱导. 东北林业大学学报, 52 (5):49-55.
[30]	Talluri R S. 2011. Gametes with somatic chromosome number and their significance in interspecific hybridization in Fuchsia. Biologia Plantarum, 55 (3):596-600.
[31]	Tian M, Zhang Y, Liu Y, Kang X, Zhang P. 2018. High temperature exposure did not affect induced 2n pollen viability in Populus. Plant,Cell Environment, 41 (6):1383-1393.
[32]	Tränkner C, Günther K, Sahr P, Engel F, Hohe A. 2020. Targeted generation of polyploids in Hydrangea macrophylla through cross-based breeding. BMC Genetics, 21 (1):147.
[33]	Wan Y, Duncan D R, Rayburn A L, Petolino J F, Widholm J M. 1991. The use of antimicrotubule herbicides for the production of doubled haploid plants from anther-derived maize callus. Theoretical and Applied Genetics, 81 (2):205-211.

花蕾直径/mm Flower bud diameter	花药宽/mm Anther width	花药长/mm Anther length	花序状态 Morphology of inflorescence	减数分裂进程 Meiosis stage
1.30 ~ 1.70	0.30 ~ 0.60	0.50 ~ 0.60	不育花未张开、花梗伸长、小叶与花序有一定夹角 Decorative florets closed，pedicel elongating，bracts angled	小孢子母细胞 Pollen mother cell
1.70 ~ 1.90	0.50 ~ 0.70	0.60 ~ 0.70	不育花未张开、花梗伸长并张开、小叶基本打开 Decorative florets closed，pedicel spreading，bracts open	细线期—粗线期 Leptotene-pachytene
1.90 ~ 2.00	0.60 ~ 0.08	0.65 ~ 0.80	不育花膨大、花梗伸长、花蕾分散、小叶完全打开 Decorative florets swollen，pedicel elongate，buds loose，bracts fully open	终变期 Diakinesis
2.00 ~ 2.10	0.60 ~ 0.85	0.80 ~ 0.85	不育花开时打开、花梗进一步伸长、花蕾进一步分散Decorative florets opening，pedicel further elongate，buds more dispersed	中期Ⅰ—前Ⅱ Metaphase I-ProphaseⅡ
2.20 ~ 2.30	0.70 ~ 0.85	0.85 ~ 0.95	不育花进一步打开、花梗和花蕾进一步分散 Decorative florets fully open，pedicel and buds widely spaced	前Ⅱ—四分体时期 Prophase Ⅱ-Tetrad
> 2.30	0.85 ~ 0.90	0.95 ~ 1.00	不育花张开、小叶成长为正常叶片大小 Decorative florets open，bracts reach mature leaf size	成熟配子 Mature gametophyte

花蕾直径/mm Flower bud diameter	花药宽/mm Anther width	花药长/mm Anther length	花序状态 Morphology of inflorescence	减数分裂进程 Meiosis stage
1.30 ~ 1.70	0.30 ~ 0.60	0.50 ~ 0.60	不育花未张开、花梗伸长、小叶与花序有一定夹角 Decorative florets closed，pedicel elongating，bracts angled	小孢子母细胞 Pollen mother cell
1.70 ~ 1.90	0.50 ~ 0.70	0.60 ~ 0.70	不育花未张开、花梗伸长并张开、小叶基本打开 Decorative florets closed，pedicel spreading，bracts open	细线期—粗线期 Leptotene-pachytene
1.90 ~ 2.00	0.60 ~ 0.08	0.65 ~ 0.80	不育花膨大、花梗伸长、花蕾分散、小叶完全打开 Decorative florets swollen，pedicel elongate，buds loose，bracts fully open	终变期 Diakinesis
2.00 ~ 2.10	0.60 ~ 0.85	0.80 ~ 0.85	不育花开时打开、花梗进一步伸长、花蕾进一步分散Decorative florets opening，pedicel further elongate，buds more dispersed	中期Ⅰ—前Ⅱ Metaphase I-ProphaseⅡ
2.20 ~ 2.30	0.70 ~ 0.85	0.85 ~ 0.95	不育花进一步打开、花梗和花蕾进一步分散 Decorative florets fully open，pedicel and buds widely spaced	前Ⅱ—四分体时期 Prophase Ⅱ-Tetrad
> 2.30	0.85 ~ 0.90	0.95 ~ 1.00	不育花张开、小叶成长为正常叶片大小 Decorative florets open，bracts reach mature leaf size	成熟配子 Mature gametophyte

减数分裂时期 Meiotic stage	处理浓度（+ 2% DMSO）/% Treatment concentration	处理时间/h Treatment time	2n花粉诱导率/% Induction rate of 2n pollen
小孢子母细胞 Pollen mother cell	0.05	24	27.34 ± 1.66
		48	47.31 ± 7.11
		72	40.94 ± 1.65
	0.10	24	42.06 ± 1.93
		48	65.55 ± 6.40
		72	40.33 ± 7.42
	0.15	24	33.26 ± 9.05
		48	36.46 ± 2.45
		72	-
终变期—中期Ⅰ Diakinesis-metaphase Ⅰ	0.05	24	34.29 ± 7.50
		48	31.66 ± 5.44
		72	45.23 ± 11.81
	0.10	24	40.73 ± 3.05
		48	39.12 ± 5.42
		72	31.18 ± 9.27
	0.15	24	35.19 ± 5.76
		48	29.84 ± 10.60
		72	-

减数分裂时期 Meiotic stage	处理浓度（+ 2% DMSO）/% Treatment concentration	处理时间/h Treatment time	2n花粉诱导率/% Induction rate of 2n pollen
小孢子母细胞 Pollen mother cell	0.05	24	27.34 ± 1.66
		48	47.31 ± 7.11
		72	40.94 ± 1.65
	0.10	24	42.06 ± 1.93
		48	65.55 ± 6.40
		72	40.33 ± 7.42
	0.15	24	33.26 ± 9.05
		48	36.46 ± 2.45
		72	-
终变期—中期Ⅰ Diakinesis-metaphase Ⅰ	0.05	24	34.29 ± 7.50
		48	31.66 ± 5.44
		72	45.23 ± 11.81
	0.10	24	40.73 ± 3.05
		48	39.12 ± 5.42
		72	31.18 ± 9.27
	0.15	24	35.19 ± 5.76
		48	29.84 ± 10.60
		72	-

变异来源 Source of variation	df	F	P
减数分裂时期 Meiotic stage	1	8.61^*	0.006
处理浓度Treatment concentration	2	50.60^**	< 0.001
处理时间Treatment time	2	26.24^**	< 0.001
减数分裂时期 × 处理浓度Meiotic stage × treatment concentration	2	4.25^*	0.022
减数分裂时期 × 处理时间Meiotic stage × treatment time	2	10.63^**	< 0.001
处理浓度 × 处理时间Treatment concentration × treatment time	4	22.63^**	< 0.001
减数分裂 × 处理浓度 × 处理时间Meiotic stage × treatment concentration × treatment time	4	0.85	0.503

High Temperature and Trifluralin Induce 2n Pollen in Hydrangea macrophylla

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不同浓度下氟乐灵处理 Treated with different concentrations of trifluralin treatment		不同时间下氟乐灵处理 Treated with different times of trifluralin treatment
处理浓度/% Treatment concentration	平均2n花粉诱导率/% Average induction rate of 2n pollen	处理时间/h Treatment time	平均2n花粉诱导率/% Average induction rate of 2n pollen
0.05	37.80 ± 9.42 b	24	35.48 ± 6.87 b
0.10	43.16 ± 12.07 a	48	41.65 ± 13.64 a
0.15	22.45 ± 17.28 c	72	26.28 ± 20.43 c

减数分裂时期 Meiotic stage	处理温度/℃ Treatment temperature	处理时间/h Treatment time	2n花粉诱导率/% Induction rates of 2n pollen
细线期—粗线期 Leptotene-pachytene	38	2	3.07 ± 0.93
		4	4.83 ± 2.49
		6	5.12 ± 0.75
	40	2	5.19 ± 0.62
		4	5.75 ± 1.29
		6	7.76 ± 1.12
	42	2	5.20 ± 0.25
		4	1.14 ± 0.91
		6	-
终变期—中期Ⅰ Diakinesis-metaphase Ⅰ	38	2	5.46 ± 1.47
		4	13.96 ± 1.91
		6	17.06 ± 3.53
	40	2	12.12 ± 1.47
		4	14.44 ± 3.92
		6	21.41 ± 2.83
	42	2	8.13 ± 1.43
		4	10.04 ± 0.71
		6	-

变异来源 Source of variation	df	F	P
减数分裂时期 Meiotic stage	1	205.70^**	< 0.001
处理温度Treatment temperature	2	64.97^**	< 0.001
处理时间Treatment time	2	7.91^**	0.001
减数分裂时期 × 处理温度Meiotic stage × treatment temperature	2	14.13^**	< 0.001
减数分裂时期 × 处理时间Meiotic stage × treatment time	2	9.25^**	< 0.001
处理温度 × 处理时间Treatment temperature × treatment time	4	26.11^**	< 0.001
减数分裂 × 处理温度 × 处理时间Meiotic stage × treatment temperature × treatment time	4	9.78^**	< 0.001

不同温度下高温处理 Treated with different temperature of high temperature treatment		不同时间下高温处理 Treated with different times of high temperature treatment
处理温度/℃ Treatment temperature	平均2n花粉诱导率/% Average induction rate of 2n pollen	处理时间/h Treatment time	平均2n花粉诱导率/% Average induction rate of 2n pollen
38	8.25 ± 5.69 b	2	6.52 ± 3.13 b
40	11.11 ± 6.12 a	4	8.36 ± 5.33 a
42	4.29 ± 3.95 c	6	7.88 ± 5.94 a

[1]	ZHANG Yu, GENG Xining, PENG Chaofeng, LI Meiyu, QI Shuaizheng, YANG Yingjun, CHENG Shiping. Pollen Chromosome Doubling of Ehretia macrophylla Induced by High Temperature [J]. Acta Horticulturae Sinica, 2025, 52(10): 2666-2676.
[2]	YANG Jun, KONG Yu, LIU Qunlu, YE Kang, QIN Jun. Effects of Shading on the Variation of Sepal Color and Pigment Composition in Hydrangea macrophylla‘Hanatemari’ [J]. Acta Horticulturae Sinica, 2023, 50(7): 1467-1481.
[3]	CHU Li-Hong, PENG Jia-Jia, WANG Zhao, ZHAO Kai, YUAN Su-Xia, MING Jun, LIU Chun. Studies on Polyploid Induction in Anthurium andraeanum Using Oryzalin，Trifluralin and Colchicine [J]. ACTA HORTICULTURAE SINICA, 2014, 41(11): 2275-2280.