Molecular Cytogenetic Identification of a Wheat (Triticum aestivum L.)-Barley(Hordeum vulgare ssp. distichon Hsü.) Dwarf Introgression Line WB29 and Its Dwarf Traits Genetic Analysis
Abstract：The new germplasm WB29 with an average height of 57 cm was a dwarf line of wheat (Triticum aestivum L.) which was selected from the derived lines after crossing and backcrossing multi-generational between common wheat 7182 and two-rowed barley (Hordeum vulgare ssp. distichon Hsü.). Cytology observation, plants height investigation, gibberellic acid reaction and molecular markers technology were used to identify this material and its heredity of dwarf traits. Cytology identification showed that the chromosome number of root-tip cell was 2n=42, no obvious hybridization signal was observed in genomic in situ hybridization (GISH) with two-rowed barley genomic DNA acted as a probe. Specific sequence-tagged site (STS) markers of barley ABG459 (2HS) could amplify specific bands of barley from WB29. Ten pairs of specific molecular markers for wheat dwarfing genes (reduced height gene, Rht) were applied to identify two-rowed barley, 7182, WB29 and Chinese spring and the result showed that 7182 and WB29 had Rht-D1b while none of the 10 Rht were detected in barley and Chinese spring. BC1F1 and F2 population were derived from crossing between WB29 and Chinese, and the statistical analysis of these two derived population indicated the dwarf traits of WB29 was controlled by one pair of incomplete dominance gene. Gibberellic acid reaction pointed that WB29 was a gibberellic-insensitive type of wheat. STS of barley ABG459 (2HS) was used to evaluate F2 population and 343 plants were found to have the barley 2HS chromosome. Wheat dwarfing gene Rht-D1b in F2 population was also detected and the result showed that 322 plants contained Rht-D1b. Linkage analysis showed that STS marker ABG459 was closely linked to the main dwarfing gene of WB29 at a genetic distance of 7.0 cM, while Rht-D1b was not linked to this main dwarfing gene. The result above had defined that WB29 did contain the barley 2HS chromosome and the dwarfing gene of WB29 which had played an important role in reducing the plant height may be located right in the barley 2HS chromosome sequence or near its insertion site. The conclusion of this research had fixed that WB29 was a wheat-barley hybrids which not only enriched the germplasm resources of wheat-barley hybrids, but also enriched the dwarf germplasm and had a certain significance in expending the application field of dwarfing genes since the main dwarfing gene of WB29 may be derived from barley and it may be a new dwarf resource.
陈新宏, 王小利, 刘淑会, 等. 2004. 普通小麦×大麦杂交后代中间材料的GISH及PAGE鉴定[J]. 遗传学报, 31(7): 730-734. (Chen X H, Wang X L, Liu S H, et al. 2004. Identification of wheat and barley hybrid offspring by GISH and PAGE[J]. Acta Genetica Sinica, 31(7): 730-734.)郭保宏, 宋春华, 贾继增. 1997. 我国小麦品种的 Rht1、Rht2矮秆基因鉴定及分布研究[J]. 中国农业科学, 30(5): 56-60. (Guo B H, Song C H, Jia J Z. 1997. Identification and distribution of Rht1 and Rht2 dwarf genes in Chinese elite dwarf wheat varieties[J]. Scientia Agricultura Sinica, 30(5): 56-60.)敬樊, 王亮明, 武军, 等. 2015. 利用甲基磺酸乙酯(EMS)诱导小麦-华山新麦草染色体易位的研究[J]. 农业生物技术学报, 23(5): 561-570. (Jing F, Wang L M, Wu J, et al. 2015. Study on inducing chromosome translocation of wheat psathyrostachys huashanica using EMS[J]. Journal of Agricultural Biotechnology, 23(5):561-570.) 刘文轩, 阎新甫, 王胜军, 等. 1992. 大麦DNA导入小麦产生的性状变异及其遗传表现[J]. 河南农业大学学报, 26(2):169-173. (Liu W X, Yan X F, Wang S J, et al. Variation of characters in wheat after introduction of barley DNA and their genetic expression[J]. Acta Agriculturae Universitatis Henanensis, 26(2): 169-173.)莫惠栋. 1984. 最大似然法及其应用[J]. 遗传, 6(5): 42-48. (Mo H D. 1984. The maximum likelihood method and its application[J]. Hereditas, 6(5): 42-48.)孙树贵, 刘淑会, 鲁敏, 等. 2014. 小麦-大麦矮秆易位系T2DL?2HS的鉴定[J]. 西北植物学报, 34(1): 21-25. (Sun S G, Liu S H, Lu M, et al. 2014. Identification of a dwarf translocation line T2DL?2HS from wheat-barley descendants[J]. Acta Botanica Boreali-Occidentalia Sinica, 34(1): 21-25.)唐娜, 姜莹, 何蓓如, 等. 2009. 赤霉素敏感性不同矮秆基因对小麦胚芽鞘长度和株高的效应[J]. 42(11): 3774-3784. (Tang N, Jiang Y, He B R, et al. 2009. Effects of dwarfing genes of Rht-B1b, Rht-D1b and Rht8 with different response to GA3 on coleoptile length and plant height of wheat[J]. Scientia Agricultura Sinica, 2009, 42(11): 3774-3784.)汪丽泉, 朱汉如, 梁竹青, 等. 1982. 中国春小麦(6x)×苏联球茎大麦(4x)属间杂交的研究初报[J]. 作物学报, 8(2): 95-102.(Wang L Q, Zhu H R, Liang Z Q, et al. 1982. A preliminary study on intergeneric crosses between wheat variety AR.Chinese spring(6×) and Hordeum bulbosum(4×)[J]. Acta Agronomica Sinica, 8(2): 95-102.)杨松杰, 张晓科, 何中虎, 等. 2006. 用 STS 标记检测矮秆基因 Rht-B1b和 Rht-D1b在中国小麦中的分布[J]. 中国农业科学, 39(8): 1680-1688. (Yang S J, Zhang X K, He Z H, et al. 2006. Distribution of dwarfing genes Rht-B1b and Rht-D1b in Chinese bread wheats detected by STS marker[J]. Scientia Agricultura Sinica, 39(8): 1680-1688.)原亚萍, 陈孝, 肖世和, 等. 2000. 应用基因组原位杂交及RFLP标记鉴定小麦中的大麦染色体[J]. 遗传学报, 27(12): 1080-1083. (Yuan Y P, Chen X, Xiao S H, et al. 2000. Identification of the barley chromosome in wheat by GISH and RFLP[J]. Acta Genetica Sinica, 27(12): 1080-1083.)周阳, 何中虎, 张改生, 等. 2003. 用微卫星标记鉴定中国小麦品种中 Rht8 矮秆基因的分布[J]. 作物学报, 29(6):810-814.(Zhou Y, He Z H, Zhang G S, et al. 2003. Rht8 dwarf gene distribution in Chinese wheats identified by microsatellite marker[J]. Acta Agronomica Sinica, 29(6): 810-814.)周之杭, 杜荣骞, 安祝平, 等. 1979. 大麦和小麦杂种胚的离体培养及杂种幼苗的形态和染色体观察[J]. 遗传学报, 6(3): 343-348.(Zhou Z H, Du R Q, An Z P, et al. 1979. Embryonic culture in vitro and observation on the seeding morphology and the chromosomes of the hybrids of barley×wheat. Acta Genet Sinica, 6(3): 343-348.) 昝凯, 孙树贵, 刘淑会, 等. 2015. 小麦-大麦大穗大粒渐渗系WB13的分子细胞学鉴定[J]. 农业生物技术学报, 23(1): 61-70. (Zan K, Sun S G, Liu S H, et al. 2015. Molecular cytogenetic identification of a large spike and grain type Wheat(Triticum aestivum)- Barley(Hordeum vulgare ssp. distichon) hybrids introgression line WB13[J]. Journal of Agricultural Biotechnology, 23(1): 61-70.)詹海仙, 畅志坚, 李光蓉, 等. 2014. 小麦-中间偃麦草抗条锈病渗入系的分子细胞学鉴定[J]. 农业生物技术学报, 22(7): 841-845. (Zhan H X, Chang Z J, Li G R, et al. 2014. Cytogenetic and molecular identification of a wheat(Triticum aestivum)-Thinopyrum intermedium introgression line with resistance to stripe rust[J]. Journal of Agricultural Biotechnology, 22(7): 841-845.)Allan R E, Vogel O A, Craddock J C. 1959. Comparative response to gibberellic acid of dwarf, semidwarf, and standard short and tall winter wheat varieties[J]. Agromomy Journal, 51(12): 737-740.Blake T K, Kadyrzhanova D, Shepherd K W, et al. 1996. STS-PCR markers appropriate for wheat-barley introgression [J]. Theoretical and Applied Genetics, 93: 826-832B?rner A, Plaschke J, Korzun V, et al. 1996. The relationships between the dwarfing genes of wheat and rye[J]. Euphytica, 89(1): 69-7Ellis M H ,Spielmeyer W, Gale K R, et al. 2002. “Perfect” markers for the Rht-B1b and Rht-D1b dwarfing genes in wheat[J]. Theoretical and Applied Genetics, 105(6-7): 1038-1042.Ellis M H, Rebetzke G J, Azanza F, et al. 2005. Molecular mapping of gibberellin-responsive dwarfing genes in bread wheat[J]. Theoretical and Applied Genetics, 111(3): 423-430.Fedak G. 1977. Increased homoeologeous chromosome pairing in Hordeum vulgare×Triticum aestivum hybrids[J]. Nature, 266(5602): 529-530.Gale M D, Law C N, Marshall G A, et al. 1975. The genetic control of gibberellic acid insensitivity and coleoptile length in a "dwarf" wheat[J]. Heredity, 34(3): 393-399. Korzun V, R?der M S, Ganal M W, et al. 1998. Genetic analysis of the dwarfing gene (Rht8) in wheat. Part I. Molecular mapping of Rht8 on the short arm of chromosome 2D of bread wheat (Triticum aestivum L.)[J]. Theoretical and Applied Genetics, 96(8): 1104-1109.Kruse A. 1973. Hordeum × Triticum hybrids[J]. Hereditas, 73(1): 157–161.Lukaszewski A J, Lapinski B, Rybka K. 2005. Limitations of in situ hybridization with total genomic DNA in routine screening for alien introgressions in wheat[J]. Cytogenetic and Genome Research, 2005, 109(1-3): 373-377.Murray M G, Thompson W F. 1980. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 8(19): 4321-4326.Qi L, Friebe B, Zhang P, et al. 2007. Homoeologous recombination, chromosome engineering and crop improvement[J]. Chromosome Research, 15(1): 3-19.Tang S, Li Z, Jia X, et al. 2000. Genomic in situ hybridization(GISH) analyses of Thinopyrum intermedium, its partial amphiploid Zhong5, and disease-resistant derivatives in wheat[J]. Theoretical and Applied Genetics, 100(3-4): 344-352.Worland A J, Korzun V, R?der M S, et al. 1998. Genetic analysis of the dwar?ng gene Rht8 in wheat. Part II. The distribution and adaptive signi?cance of allelic variants at the Rht8 locus of wheat as revealed by microsatellite screening[J]. Theoretical and Applied Genetics, 96(8): 1110-1120.Wu J, Yang X M, Wang H, et al. 2006. The introgression of chromosome 6P specifying for increased numbers of florets and kernels from Agropyron cristatum into wheat [J]. Theoretical and Applied Genetics, 114(1): 13-20.Zhao J X, Du W L, Wu J, et al. 2013. Development and identification of a wheat-Leymus mollis multiple alien substitution line[J]. Euphytica, 190(1): 45-52.
