小麦与玉米杂交产生小麦单倍体与双单倍体的稳定性

Transcription

1 作物学报 ACTA AGRONOMICA SINICA 2013, 39(12): ISSN ; CODEN TSHPA9 DOI: /SP.J 小麦与玉米杂交产生小麦单倍体与双单倍体的稳定性 * 陈新民 王凤菊李思敏张文祥, :, 13 F 1,, 23.9%, 6.8, 3 ; Dicamba 21.5%, 2,4-D (21.1%), B %~88.3%, 82.0%; 1/2 MS 70.0%~86.0%, 76.6%; 67.6% 8.6%,, : ; ; ; Stable Production of Wheat Haploid and Doubled Haploid by Wheat Maize Cross CHEN Xin-Min *, WANG Feng-Ju, LI Si-Min, and ZHANG Wen-Xiang Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing , China Abstract: One of the most effective ways to produce wheat haploid is the method of crossing wheat with maize. However, the problem of unstable frequencies in haploid and doubled haploid production affects its application. In this study, 13 wheat F 1 hybrids crossed with maize to explore the effects of wheat growth condition, hormone treatment, culture medium, and strong seedling treatment on production of haploid and doubled haploid. The frequency of embryo formation was 23.9% and the number of embryos per spike was 6.8 for the wheat plants grown in filed and then detached before pollination and hydroponically cultured after pollination. The efficiency of this method was three times higher than that of the method with plants transplanted into pot after green recovering in early spring and then grown in the unheated greenhouse condition. Embryo formation frequencies were obviously different among wheat crosses. There was no significant difference in the frequency of embryo formation between treatments of dipping spike with Dicamba (21.5%) and 2,4-D (21.1%). However, wheat cross had significant effect on embryo formation. The mean frequency of embryo germination on B 5 medium was 82.0%, ranging from 70.9% to 88.3%, whereas averaged frequency of embryo germination was 76.6% on 1/2 MS medium, ranging from 70.0% to 86.0%. There was no significant difference between the two culture media in embryo germination. The efficiency of doubled haploid production for the seedlings treated with culture medium and transplanted into pot was 67.6% and 8.6%, respectively. Most of seedlings transplanted into pot had no tiller and grew weakly; this is the reason for low frequency of survival plant and very low frequency of doubling after colchicine treatment. Keywords: Wheat maize cross; Haploid; Hormone treatment; Culture medium,,, 21,,, DH (doubled haploid) [1-2] 3:, Kr, [3-4] ;,, [2,5-7],, (2011BAD35B03)(948 ) (2011-G3) * (Corresponding author):, chenxinmin@caas.cn Received( ): ; Accepted( ): ; Published online(): URL:

2 [8], ;,,,,,,,, 3~4, ; DHQTL, Laurie [9], [10-11], 1997 DH, , 23 DH, 2009 DH 1/3 [12] 1992,, [13],,,, DH 1 材料与方法 F 1 13 ( 1), CRC1 (Cereal Research Centre, Agriculture and Agri- Food Canada) 1.2,,,, 9:00 10:00, (20~23 ), 1/2 MS ( )+8 ml L g L 1, 16:00 17:00, : 18~20, 60%~70%, 16 h /8 h 1 d, 100 mg L 1 2,4-D ; 2 d, 2 d,, 16:00 17:00 ( ) 1 d 100 mg L 1 2,4-D, 2 d, 2,4-D Dicamba, 100 mg L 1, ~16 d,, 70% 30 s, 0.1% 10 min, 3~4,, 1/2 MS ( 2%, 7.5 g L 1 ) B 5 ( 2%, 10 g L 1 ) 4 ( ) 2~3 d, 2 d, 20~23 16 h /8 h 1.4 3~5 cm, (1/2 MS+ 0.5 mg L 1 + IAA 0.2 mg L 1 ), 11~15, 16 h 2 ;, 60% 2, 16 (N P K= ) 3 1~3, 2~6 1 ( 16 h d 1 ), 3~4 d, 1.5 ( 1~2 cm), 0.2% +DMSO ( 10 ml 1 ), 3~4 h, 3~4 h, 2,, 18~22,,, 1.6, 3,,, 3,, 6, 6~10 ( ), (%)= / 结果与分析 2.1, 7 d,, ( 1) 6 5.8%~10.4%, 7.8%; 1.0~2.5, %~29.6%, 23.9%; 3.6~8.5, 6.8,, Dicamba 2,4-D ( 2) Dicamba %~27.8%, 21.5%; 2,4-D

3 12 : 2249 Table 1 Cross 表 1 不同生长条件下小麦 玉米得胚率 Embryo Formation by wheat maize method under different growing conditions No. of spikes pollinated Plants transplanted into pot during green recovering in early spring No. of florets pollinated No. of embryos formed Frequency of embryos formed (%) No. of embryos per spike 175/CA0996 Zhongmai 175/CA CA0816/ 15 CA0816/Shimai CA0816/ 5067 CA0816/Bao CA0415/CA /CA9722 Lunxuan 987/CA / 175 Lunxuan 987/Zhongmai Mean Plants grown in field and then detached and hydroponically cultured after emasculation CA0415/CA CA0996/ 175 CA0996/Zhongmai CA0998/CA /CA0477 Lunxuan 987/CA / 175 Lunxuan 987/Zhongmai CA0629/CA / 15 Yangmai 16/Yangmai / 895 Yangmai 16/Zhongmai Mean Cross Table mg L 1 Dicamba Treated with 100 mg L 1 Dicamba 表 2 不同激素处理效应 Effect of different hormone treatments No. of florets pollinated No. of caryopses Frequency of caryopses (%) No. of embryos formed Frequency of embryos formed (%) CA0415/CA CA0996/ 175 CA0996/Zhongmai CA0998/CA /CA0477 Lunxuan 987/CA Mean mg L 1 2,4-D Treated with 100 mg L 1 2,4-D CA0415/CA CA0996/ 175 CA0996/Zhongmai CA0998/CA /CA0477 Lunxuan 987/CA Mean %~28.3%, 21.1% Dicamba (82.4%) 2,4-D (58.5%), CA0998/CA0816 (28.1%), CA0996/ 175 (24.2%), 987/CA0477 (20.5%) CA0415/CA0391 (12.4%) 2.3 B5 82.0%, 70.9%~88.3%; 1/2 MS 70.0%~86.0%, 76.6%; ( 3), CA0998/CA0816 (87.2%), 16/ 15 (73.2%)987/CA0477 (73.3%), CA0415/CA0391 (79.2%) CA0996/ 175 (81.9%) 16/ 895 (81.3%)

4 , 92.0% 67.6%, 46.2% 8.6%, ( 4),,,, Cross B 5 B 5 medium Table 3 表 3 不同培养基胚培养效果 Effect of different culture media on embryo culture No. of embryos cultured No. of germinated embryos Frequency of germination (%) CA0415/CA CA0996/ 175 CA0996/Zhongmai CA0998/CA /CA0477 Lunxuan 987/CA / 15 Yangmai 16/Yangmai / 895 Yangmai 16/Zhongmai Mean /2 MS 1/2 MS medium CA0415/CA CA0996/ 175 CA0996/Zhongmai CA0998/CA /CA0477 Lunxuan 987/CA / 15 Yangmai 16/Yangmai / 895 Yangmai 16/Zhongmai Mean 76.6 Table 4 Method for seedling treatment Transfer plantlets to pot for strong seedling Transfer plantlets to medium for strong seedling 表 4 不同壮苗处理方法加倍效果 Effect of different strong seedling treatments on doubling No. of plants treated with colechicine No. of survived plants Survival frequency (%) Plants with seeds Efficiency of doubling (%) 讨论,, [1,4],,, [9], 2,4-D, Dicamba [14-15] Knox [16] Dicamba 2,4-D, Dicamba 2,4-D, 100 mg L 1, Dicamba 2,4-D,, Dicamba,,, Dicamba 2,4-D, 2,4-D, [17], (29.9%) (3.6%),,, [18-21],,, 3, 2

5 12 : 2251, 2 987/ 175 CA0996/ 175 (), 27.2% 24.4%, 10.4% 7.2%, 2011 ( ),,,,, ;,,,,,,,,,, 1/2 MS, Difco Orchid B 5 [2,22] B 5 1/2 MS, 5.4, [21], [23] 6 7, 4 3~4,,, 2 DH DH,, ,,,,,,,, (), 6 7,,, 12 1 DH,,, DH,, 3, 1000 DH, % 67%, 268 DH 7~8, 15 m 2, 20~30 m 2, 15 m 2, (Cereal Research Centre, Agriculture and Agri-Food Canada) DH, 3, 6~7,, DH References [1] Laurie A D, Bennett M D. Wheat maize hybridization. Can J Genet Cytol, 1986, 28: [2] Laurie A D, Reymondie S. High frequencies of fertilization and haploid seeding production in crosses between commercial hexaploid wheat varieties and maize. Plant Breed, 1991, 106: [3] Inagaki M, Tahir M. Comparison of haploid production frequency in wheat varieties crossed with Hordeum bulbosum L. and maize. Jpn J Breed, 1990, 40: [4] Laurie D A, Bennett M D. The effect of the crossability loci Kr1 and Kr2 on fertilization frequency in hexaploid wheat maize crosses. Theor Appl Genet, 1987, 73: [5] Chen X-M(), Lai G-X(), Chen X( ), Zhou J-F(), Liu J-X(), Sun F-H(). Difference of haploid production in crosses between different F 1 and maize. Acta Agron Sin ( ), 1996, 22(4): (in Chinese with English abstract) [6] Mujeeb-Kazi A, Cortes A, Riera-Lizarazu O, Faridi N I, Delgado R. Polyhaploidy in the Triticeae madited by crosses of Triticum species with Zea mays and Trip sacumdatyloides. In: 8th Int Wheat Genet Symp. Beijing: China Agricultural Science and Technology Press, pp [7] Suenaga K, Nakajima K. Efficient production of haploid wheat (Triticum aestivum) through crosses between Japanese wheat and maize (Zea mays). Plant Cell Rep, 1989, 8: [8] Kisan N S, Nkongolo K K, Quick J S, Johnson D L. Production of doubled haploids by anther culture and wheat maize method in a wheat breeding programme. Plant Breed, 1993, 110: [9] Laurie A D, Bennett M D. The production of haploid wheat plants from wheat maize crosses. Theor Appl Genet, 1988, 76: [10] Chen X-M(), Chen X( ). Progress on haploid and doubled haploid production by wheat maize method. Tritical Crops ( ), 1998, 18(3): 1 4 (in Chinese) [11] Cai H( ), Ma C-X(), Si H-Q(), Qiao Y-Q( ), Lu W-Z(). Advance on producing double haploid of wheat by wide hybridization between wheat and maize. J Triticeae Crops (), 2006, 26(4): (in Chinese with English abstract) [12] DePauw R M, Knox R E, Humphreys D G. New breeding tools impact Canadian commercial farmer fields. Czech J Genet Plant Breed, 2011, 47: S28 S34 [13] Chen X-M(), He Z-H(), Liu C-L(), Wang D-S(), Zhang Y( ). New wheat variety Zhongmai 533 developed by wheat maize cross. J Triticeae Crops ( ), 2011, 31(3): (in Chinese with English abstract) [14] Matzk F, Mahn A. Improved techniques for haploid production in wheat using chromosome elimination. Plant Breed, 1994, 113: [15] Campbell A W, Grin W B, Conner A J, Rowarth J S, Burritt D J. The effects of temperature and light intensity on embryo numbers in wheat doubled haploid production through wheat maize crosses. Ann Bot, 1998, 82: [16] Knox R E, Clarke J M, DePauw R M. Dicamba and growth con-

6 dition effects on doubled haploid production in durum wheat crossed with maize. Plant Breed, 2000, 119: [17] Chen X-M(), Chen X( ), Li X-Y(). Study on increasing the frequency of haploid production in wheat maize crosses. In: Symposium on Crop Breeding ( ). Beijing: China Agricultural Science and Technology Press, pp (in Chinese with English abstract) [18] Gu J( ), Liu K( ), Li S-X(), Tian Y-X(), Yang H-X(), Yang M-J(). Study on the in vitro culture of cut plant in wheat haploid embryo induction by wheat maize cross. J Triticeae Crops (), 2008, 28(1) : 1 5 (in Chinese with English abstract) [19] Cai H( ), Ma C-X(), Si H-Q(), Qiao Y-Q( ), Lu W-Z(). Study on increasing frequency of haploid embryo production through wheat maize. J Triticeae Crops ( ), 2005, 25(6): (in Chinese with English abstract) [20] Inagaki M. Use of pollen storage and detached tiller culture in wheat polyploidy production through wide cross. Cereal Res Commun, 1997, 25: 7 13 [21] Cherkaoui S, Lamsaouri O, Chlyah A, Chlya H H. Durum wheat maize crosses for haploid wheat production: influence of parental genotypes and various experimental factors. Plant Breed, 2000, 119: [22] Lefebre D, Devaux P. Doubled haploids of wheat from wheat maize crosses: genotype influence, fertility and inheritance of the 1BL/1RS chromosome. Theor Appl Genet, 1996, 93: [23] Chen X M, Zhang W X, Cui S L, Chen X. Study on chromosome doubling for haploid produced by wheat maize crossing. Agric Sci China, 2002, 1: