小麦花药培养最佳取样时期穗部形态特征鉴定与高培养力基因型筛选

doi:10.3724/SP.J.1006.2025.51023

Abstract

Abstract:

Haploid breeding based on anther culture is an important method for the efficient selection of wheat varieties. However, the efficiency of anther culture varies significantly among different wheat genotypes, which limits its broader application in wheat breeding. To date, a few wheat lines with high anther culture efficiency—such as Shi4185, H307, and Zhoumai16—have been identified. Nevertheless, there remains a lack of sufficient high-performing materials to support large-scale application of this technique. In this study, we systematically evaluated anther culture-related traits in 94 wheat varieties grown under greenhouse conditions. The optimal anther sampling stage for these varieties was determined to occur when the distance between the tip of the developing spike and the leaf auricle ranged from -5 cm to 2 cm, with the highest sampling frequency observed at -2 cm. Notably, spike morphological characteristics associated with the optimal sampling period differed between plants grown under greenhouse and field conditions. Among the 94 greenhouse-grown varieties, the callus induction rate, green shoot differentiation rate, albino shoot differentiation rate, and green plantlet production rate during anther culture ranged from 0-15%, 0-100%, 0-60%, and 0-22.95%, respectively. The callus induction rate showed highly significant positive correlations with the green shoot differentiation rate, albino shoot differentiation rate, and green plantlet production rate. Moreover, the callus induction rate exhibited consistent performance across both greenhouse and field conditions. Of the 94 varieties tested, fourteen exhibited green plantlet production rates exceeding 1%, and six varieties met the criteria for high anther culture efficiency while also demonstrating excellent agronomic traits. These varieties represent valuable germplasm resources for haploid breeding in wheat. This study provides practical techniques and genetic materials to support the advancement of wheat haploid breeding programs.

Key words: wheat, double haploid, anther culture, high anther culture ability, breeding application

LU Xiang-Qian, FU Yu-Jie, ZHAO Jun-Heng, ZHENG Nan-Nan, SUN Nan-Nan, ZHANG Guo-Ping, YE Ling-Zhen. Characterization of spike morphological traits at optimal sampling stage and screening of high-culturability genotypes in wheat anther culture[J].Acta Agronomica Sinica, 2025, 51(8): 2033-2047.

Figures/Tables 11

Table S1

Traits of anther culturing ability of 94 wheat varieties grown in greenhouse"

品种 Variety	愈伤诱导率 Callus induction rate (%)	绿芽分化率 Green shoot differentiation rate (%)	绿苗产率 Green plantlet production rate (%)	白芽分化率 Albino shoot differentiation rate (%)	取样特征 Sampling feature (cm)
淮麦20 Huaimai 20	0	0	0	0	-1
宿0663 Su 0663	0	0	0	0	-2
阜936 Fu 936	0	0	0	0	-2
宿农6号 Sunong 6	0	0	0	0	-2
皖麦33 Wanmai 33	0.97	0	0	0	-3
皖麦52 Wanmai 52	1.15	0	0	0	-3
皖麦50 Wanmai 50	0	0	0	0	-3
皖麦29 Wanmai 29	0	0	0	0	-5
皖麦53 Wanmai 53	0	0	0	0	-2
石新828 Shixin 828	0	0	0	0	0
石家庄15 Shijiazhuang 15	3.33	20.00	1.33	60.00	-1
衡7228 Heng 7228	9.76	83.33	6.50	16.67	-1
冀师02-1 Jishi 02-1	0	0	0	0	1
石新733 Shixin 733	0	0	0	0	-3
石4185 Shi 4185	14.29	55.56	15.87	22.22	0
高优503 Gaoyou 503	0	0	0	0	-3
藁城8901 Gaocheng 8901	0	0	0	0	-2
石家庄8号 Shijiazhuang 8	0.92	0	0	0	-2
石优17 Shiyou 17	0	0	0	0	0
衡观33 Hengguan 33	2.00	0	0	0	-2
偃展4110 Yanzhan 4110	1.27	0	0	0	-2
百农3217 Bainong 3217	1.54	50.00	0	0	－2
豫麦13 Yumai 13	9.52	0	0	33.33	1
中麦895 Zhongmai 895	6.48	14.29	3.70	28.57	-2
周麦19 Zhoumai 19	6.60	0	0	0	0
新麦9408 Xinmai 9408	1.41	0	0	0	-1
豫麦21 Yumai 21	0	0	0	0	0
豫麦47 Yumai 47	0	0	0	0	2
周麦25 Zhoumai 25	8.94	0	0	18.75	-3
兰考2号 Lankao 2	0	0	0	0	-2
中麦871 Zhongmai 871	0	0	0	0	-3
豫麦49 Yumai 49	2.59	0	0	0	-2
新麦9号 Xinmai 9	0	0	0	0	-3
兰考906 Lankao 906	14.00	0	0	0	-3
洛旱2号 Luohan 2	2.59	66.67	10.34	33.33	-2
豫麦57 Yumai 57	0	0	0	0	-2
豫麦18 Yumai 18	0	0	0	0	-4
周麦12 Zhoumai 12	1.20	0	0	0	-2
矮抗58 Aikang 58	0	0	0	0	-2
周麦30 Zhoumai 30	1.72	0	0	0	-2
豫麦34 Yumai 34	0	0	0	0	-3
豫麦7号 Yumai 7	4.69	0	0	16.67	-5
郑麦366 Zhengmai 366	0	0	0	0	0
中育5号 Zhongyu 5	1.36	0	0	0	-2
周麦32 Zhoumai 32	2.82	0	0	0	-2
中育9号 Zhongyu 9	9.02	0	0	27.27	-4
郑引1号 Zhengyin 1	0	0	0	0	-4
豫麦50 Yumai 50	0	0	0	0	-5
周麦23 Zhoumai 23	0	0	0	0	-1
周麦28 Zhoumai 28	1.53	0	0	33.33	-2
豫麦2号 Yumai 2	15.00	46.67	8.00	26.67	-1
周麦26 Zhoumai 26	0	0	0	0	-1
中麦875 Zhongmai 875	5.53	7.69	0.85	23.08	-2
花培5号 Huapei 5	2.36	0	0	0	-1
兰考24 Lankao 24	6.32	45.45	5.75	9.09	-3
济麦20 Jimai 20	0	0	0	0	-2
鲁麦9号 Lumai 9	0.49	0	0	0	-2
汶农5号 Wennong 5	0	0	0	0	-3
烟农18 Yannong 18	1.83	25.00	3.20	0	-5
鲁麦14 Lumai 14	2.50	40.00	2.50	0	-4
济宁16 Jining 16	0	0	0	0	-3
烟农15 Yannong 15	10.49	0	0	0	-4
鲁麦21 Lumai 21	6.06	0	0	50.00	-3
淄选2号 Zixuan 2	0.96	0	0	0	-2
泰山1号 Taishan 1	1.78	66.67	4.14	0	-2
济南17 Jinan 17	0	0	0	0	-1
汶农14 Wennong 14	7.41	0	0	10.00	-3
鲁麦8号 Lumai 8	0	0	0	0	-3
良星99 Liangxing 99	0	0	0	0	-3
鲁原502 Luyuan 502	1.50	0	0	0	-2
泰山5号 Taishan 5	1.18	100.00	2.35	0	-1
济麦21 Jimai 21	0	0	0	0	-3
鲁麦23 Lumai 23	0	0	0	0	-4
山农20 Shannong 20	4.07	0	0	0	-1
临麦4号 Linmai 4	0	0	0	0	-3
临抗12 Linkang 12	0	0	0	0	-4
碧蚂1号 Bima 1	1.69	0	0	0	-1
陕农78-59 Shaannong 78-59	1.80	0	0	0	-2
陕麦509 Shaanmai 509	3.25	25.00	1.63	0	-1
西农979-005 Xinong 979-005	2.48	0	0	0	-2
西农2000-7 Xinong 2000-7	0	0	0	0	0
陕麦354 Shaanmai 354	0.45	0	0	0	0
西农291 Xinong 291	5.00	20.00	0	0	-2
小偃54 Xiaoyan 54	0	0	0	0	0
小偃6号 Xiaoyan 6	0	0	0	0	-4
西农1376 Xinong 1376	14.75	83.33	22.95	0	-2
陕麦512 Shaanmai 512	1.95	25.00	0.98	0	-3
矮丰3号 Aifeng 3	0	0	0	0	-4
陕优225 Shaanyou 225	0	0	0	0	-2
武农148 Wunong 148	0	0	0	0	-3
陕麦229 Shaanmai 229	0	0	0	0	-5
小偃81 Xiaoyan 81	0	0	0	0	-4
陕麦94 Shaanmai 94	6.67	0	0	44.44	-2
碧蚂4号 Bima 4	1.91	0	0	0	-2

Table S1

Table 1

Fig. 1

Table S2

Table 2

Fig. 2

Fig. 3

Fig. 4

Table 3

Table 4

Table S3

Information of test varieties"

品种 Variety	育成年份 Year of cultivation	亲本 Parent
泰山5号Taishan 5	1975	(辉县红×阿勃) F₁×白欧柔(Huixianhong×Abo) F₁×Bai’ourou
藁城8901 Gaocheng 8901	1990	冀麦26×鲁麦1号Jimai 26×Lumai 1
西农1376 Xinong 1376	1992	西农84G6×比16 Xinong 84G6×Bi 16
陕优225 Shaanyou 225	1992	小偃6号×NS2761 Xiaoyan 6×NS2761
新麦9408 Xinmai 9408	1994	新麦9号×豫麦21 Xinmai 9×Yumai 21
豫麦34 Yumai 34	1994	豫麦13×鲁麦14 Yumai 13×Lumai 14
陕麦229 Shaanmai 229	1995	(小偃6号×陕7853) F₁×TB902 (Xiaoyan 6×Shaan7853) F₁×TB902
鲁麦21 Lumai 21	1996	烟中144×宝丰7228 Yanzhong 144×Baofeng 7228
鲁麦23 Lumai 23	1996	鲁麦8号×高赖小麦Lumai 8×Gaolaixiaomai
淮麦20 Huaimai 20	1997	豫麦13×鲁麦14 Yumai 13×Lumai 14
皖麦33 Wanmai 33	1997	安农2号×百农3217 Annong 2×Bainong 3217
豫麦47 Yumai 47	1997	豫麦13×鲁麦14 Yumai 13×Lumai 14
皖麦29 Wanmai 29	1997	(一粒小麦×阿夫乐尔) F0射线处理/NPFP-5442/3/ALONDRA/4/2411/矮丰3号//7679-29-2-2 (Einkorn wheat × Avrora) F0 treated by ray /NPFP-5442/3/ALONDRA/4/2411/ Aifeng 3//7679-29-2-2
石新733 Shixin 733	1998	大拇指矮×石新163 Damuzhi’ai×Shixin 163
豫麦57 Yumai 57	1999	豫麦18×80 (6)-3-3-10 Yumai 18×80 (6)-3-3-10
高优503 Gaoyou 503	2001	豫麦34×鲁麦14 Yumai 34×Lumai 14
陕麦512 Shaanmai 512	2001	陕麦150MS2×陕354 Shaanmai 150MS2×Shaan 354
兰考906 Lankao 906	2001	—
中育5号Zhongyu 5	2001	冀5418×豫西832 Ji 5418×Yuxi 832
小偃81 Xiaoyan 81	2001	小偃54×8602 Xiaoyan 54×8602
陕农78-59 Shaannong 78-59	2002	—
偃展4110 Yanzhan 4110	2003	偃展1号×豫麦18 Yanzhan 1×Yumai 18
周麦19 Zhoumai 19	2003	周麦13×豫麦49 Zhoumai 13×Yumai 49
矮抗58 Aikang 58	2003	周麦11×豫麦49 Zhoumai 11×Yumai 49
济麦20 Jimai 20	2003	济麦19×鲁麦14 Jimai 19×Lumai 14
新麦9号Xinmai 9	2003	百泉3047-3×内乡82C6 Baiquan 3047-3×Neixiang 82C6
汶农5号Wennong 5	2003	鲁麦21号×泰山5号Lumai 21×Taishan 5
中育9号Zhongyu 9	2004	豫麦21号×92R139 Yumai 21×92R139
济宁16 Jining 16	2004	(烟1934/824046) F₁×(聊83-1/2114) F₁ (Yan 1934/824046) F₁×(Liao 83-1/2114) F₁
石家庄8号 Shijiazhuang 8	2005	石4185×石新733 Shi 4185×Shixin 733
洛旱2号Luohan 2	2005	豫麦18×洛旱1号Yumai 18×Luohan 1
郑麦366 Zhengmai 366	2005	豫麦34×周麦16 Yumai 34×Zhoumai 16
良星99 Liangxing 99	2005	济麦20×鲁麦23 Jimai 20×Lumai 23
阜936 Fu 936	2005	(皖麦20×冀5418) F₁×内乡184 (Wanmai 20×Ji 5418) F₁×Neixiang 184
皖麦52 Wanmai 52	2005	郑州8329×皖麦19 Zhengzhou 8329×Wanmai 19
皖麦50 Wanmai 50	2005	豫麦29×皖麦19 Yumai 29×Wanmai 19
皖麦53 Wanmai 53	2005	豫麦29×皖麦19 Yumai 29×Wanmai 19
衡7228 Heng 7228	2005	冀5418×衡5041 Ji 5418×Heng 5041
石优17 Shiyou 17	2005	冀935352×鲁麦21 Ji 935352×Lumai 21
衡观33 Hengguan 33	2006	衡7228×石4185 Heng 7228×Shi 4185
济麦21 Jimai 21	2006	济麦19×鲁麦14 Jimai 19×Lumai 14
花培5号Huapei 5	2006	豫麦18×花4-3 Yumai 18×Hua 4-3
临麦4号Linmai 4	2006	鲁麦23号×临9015 Lumai 23×Lin 9015
西农2000-7 Xinong 2000-7	2006	(西农2611×386) F₁×(小偃22×陕354) F₁ (Xinong 2611×386) F₁×(Xiaoyan 22×Shaan 354) F₁
周麦25 Zhoumai 25	2008	周麦16×豫麦49 Zhoumai 16×Yumai 49
中麦895 Zhongmai 895	2010	中麦16×周麦18 Zhongmai 16×Zhoumai 18
周麦23 Zhoumai 23	2010	周麦16×豫麦49 Zhoumai 16×Yumai 49
鲁原502 Luyuan 502	2010	济麦22×鲁麦14 Jimai 22×Lumai 14
汶农14 Wennong 14	2010	系84139//9215/876161 Xi 84139//9215/876161
周麦26 Zhoumai 26	2012	周麦16×百农64 Zhoumai 16×Bainong 64
周麦28 Zhoumai 28	2013	周麦16×周麦22 Zhoumai 16×Zhoumai 22
石新828 Shixin 828	2013	石新163×石新422 Shixin 163×Shixin 422
周麦32 Zhoumai 32	2014	矮抗58×周麦24 Aikang 58×Zhoumai 24
中麦875 Zhongmai 875	2014	周麦16×荔垦4号Zhoumai 16×Liken 4
周麦30 Zhoumai 30	2015	周麦22×周麦18 Zhoumai 22×Zhoumai 18
山农20 Shannong 20	2015	济麦22×山农15 Jimai 22×Shannong 15
碧蚂1号Bima 1	1950s	碧玉麦×蚂蚱麦Biyumai×Mazhamai
碧蚂4号Bima 4	1950s	碧玉麦×蚂蚱麦Biyumai×Mazhamai
郑引1号Zhengyin 1	1960s	引自意大利的Strampelli from Italian strampelli
豫麦2号Yumai 2	1970s	碧蚂4号×阿夫Bima 4×Funo
泰山1号Taishan 1	1970s	碧蚂4号×欧柔Bima 4×Ourou
豫麦13 Yumai 13	1980s	郑州761×偃师4号Zhengzhou 761×Yanshi 4
豫麦21 Yumai 21	1980s	郑州761×偃师9号Zhengzhou 761×Yanshi 9
豫麦18 Yumai 18	1980s	豫麦2号×偃师4号Yumai 2×Yanshi 4
豫麦7号Yumai 7	1980s	郑州761×偃师4号 Zhengzhou 761×Yanshi 4
鲁麦9号Lumai 9	1980s	泰山1号×鲁麦5号Taishan 1×Lumai 5
鲁麦14 Lumai 14	1980s	泰山1号×鲁麦5号Taishan 1×Lumai 5
烟农15 Yannong 15	1980s	烟农73×鲁麦1号Yannong 73×Lumai 1
鲁麦8号Lumai 8	1980s	泰山1号×鲁麦5号Taishan 1×Lumai 5
小偃6号Xiaoyan 6	1980s	小偃5号×中间偃麦草Xiaoyan 5×Zhongjianyanmaicao
石4185 Shi 4185	1990s	冀麦30×鲁麦14 Jimai 30×Lumai 14
百农3217 Bainong 3217	1990s	百农791×豫麦13 Bainong 791×Yumai 13
豫麦49 Yumai 49	1990s	豫麦13×鲁麦14 Yumai 13×Lumai 14
烟农18 Yannong 18	1990s	烟农15×鲁麦14 Yannong 15×Lumai 14
济南17 Jinan 17	1990s	鲁麦14×济南13 Lumai 14×Jinan 13
石家庄15 Shijiazhuang 15	2000s	石4185×冀麦38 Shi 4185×Jimai 38
西农979-005 Xinong 979-005	2010s	西农979×周麦18 Xinong 979×Zhoumai 18
矮丰3号Aifeng 3	—	咸农39号×丰产3号Xiannong 39×Fengchan 3
周麦12 Zhoumai 12	—	周8425A×SWD73295 Zhou 8425A×SWD73295
宿0663 Su 0663	—	—
宿农6号Sunong 6	—	—
冀师02-1 Jishi 02-1	—	—
兰考2号Lankao 2	—	—
中麦871 Zhongmai 871	—	—
豫麦50 Yumai 50	—	—
兰考24 Lankao 24	—	—
淄选2号Zixuan 2	—	—
临抗12 Linkang 12	—	—
陕麦509 Shaanmai 509	—	—
陕麦354 Shaanmai 354	—	—
西农291 Xinong 291	—	—
小偃54 Xiaoyan 54	—	—
武农148 Wunong 148	—	—
陕麦94 Shaanmai 94	—	—

Table S3

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培养基种类 Medium type	基本培养基 Basic medium	外源物1 Additive 1 (mg L^-1)	外源物2 Additive 2 (mg L^-1)	蔗糖含量 Sucrose content (g L^-1)	琼脂含量 Agar content (g L^-1)
诱导培养基 Induction medium	C17	2,4D (2.0)	KT (0.5)	90.0	7.0
分化培养基 Differentiation medium	MS	NAA (1.0)	KT (1.0)	30.0	5.0
壮根培养基 Strong root medium	1/2MS	MET (3.0)	KT (1.0)	80.0	8.0

花培力指标 Index of anther culture ability	数量Number	范围 Range	均值 Mean	标准差 SD	方差 Variance
愈伤诱导率 Callus induction rate	94	0-0.150	0.023	0.037	0.001
绿芽分化率 Green shoot differentiation	49	0-1.000	0.158	0.270	0.073
白芽分化率 Albino shoot differentiation rate	49	0-0.600	0.093	0.155	0.024
绿苗产率 Green plantlet production rate	49	0-0.230	0.020	0.047	0.002

品种 Variety	株高 Plant height (cm)	穗粒数 Kernels per spike	千粒重 TKW (g)	品种特性 Varietal characteristic	参考 Reference
西农1376 Xinong 1376	73.33	35.50	42.06	早熟、高产, 遗传转化率高 Early maturity, high yield, high genetic transform rate	[19]
石4185 Shi 4185	78.00	34.00	37.00	花培育种优良亲本 Excellent parent for breeding by anther culture	农博数据, [18] Nongbo data, [18]
豫麦2号 Yumai 2	85.00	—	36.00-38.00	广适、丰产, 骨干亲本 Widely suitable, fertile, backbone parent	农博数据, [20] Nongbo data, [20]
衡7228 Heng 7228	75.00	37.00	40.00	高产、节水 High yield, water economy	农博数据 Nongbo data
兰考24 Lankao 24	73.67	46.29	41.41	—	河南荥阳测定 Measured in Xingyang, Henan province
中麦895 Zhongmai 895	70-75	35.00	48.00-52.00	高产、耐热 High yield, heat resistance	农博数据, [21] Nongbo data, [21]

Characterization of spike morphological traits at optimal sampling stage and screening of high-culturability genotypes in wheat anther culture

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品种 Variety	愈伤诱导率 Callus induction rate (%)	取样特征 Sampling feature (cm)	品种 Variety	愈伤诱导率 Callus induction rate (%)	取样特征 Sampling feature (cm)
西农1376 Xinong 1376	2.8986	-3	周麦19 Zhoumai 19	3.0769	0
石4185 Shi 4185	5.8065	0	兰考906 Lankao 906	17.0455	-2
豫麦2号Yumai 2	3.3333	-1	山农20 Shannong 20	2.4390	-1
衡7228 Heng 7228	0	-1	烟农15 Yannong 15	0	-3
中麦895 Zhongmai 895	4.7619	-1	百农3217 Bainong 3217	0.5236	-2
中麦875 Zhongmai 875	1.8692	-2	鲁麦9号Lumai 9	1.3333	-2
豫麦18 Yumai 18	0	-3	宿0663 Su 0663	2.1277	-1
偃展4110 Yanzhan 4110	6.2500	-1	济宁16 Jining 16	2.8571	-2
鲁麦21 Lumai 21	7.8431	-2	石新733 Shixin 733	1.7964	-2
郑麦366 Zhengmai 366	1.4493	0	皖麦33 Wanmai 33	0	-3
豫麦49 Yumai 49	0	-1	周麦25 Zhoumai 25	0	-3
豫麦21 Yumai 21	0	0	豫麦34 Yumai 34	0	-2
周麦32 Zhoumai 32	2.5000	-2	郑引1号Zhengyin 1	0	-3
花培5号Huapei 5	13.8298	0	周麦23 Zhoumai 23	0.6452	0
陕麦94 Shaanmai 94	3.1915	-2	周麦26 Zhoumai 26	0	-1
豫麦13 Yumai 13	0	0	石优17 Shiyou 17	2.8571	0
汶农14 Wennong 14	0	-2	衡观33 Hengguan 33	0.6667	-2
中育9号Zhongyu 9	13.1387	-2

品种 Variety	愈伤诱导率 Callus induction rate (%)	绿芽分化率 Green shoot differentiation rate (%)	绿苗产率 Green plantlet production rate (%)	白芽分化率 Albino shoot differentiation rate (%)	参考 Reference
藁城8901	0	0	0	0	本研究 This study
Gaocheng 8901	4.94	9.57	0.39	11.90	[15]
偃展4110	1.27	0	0	0	本研究 This study
Yanzhan 4110	3.97	0	0	0	[15]
洛旱2号	2.59	66.67	10.34	33.33	本研究 This study
Luohan 2	9.33	0	0	1.28	[15]
石4185	14.29	55.56	15.87	22.22	本研究 This study
Shi 4185	6.67	29.45	2.39	1.67	[15]

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