Comparison of Main Physiological Characteristics, Chloroplast Morphology and Ultra-structure of the Yellow-green Wheat (Triticum aestivum) Near-isogenic Lines
Abstract：Exploring genetic mechanism of chlorophyll biosynthesis and chloroplast development have important implication for high photosynthetic efficiency breeding in wheat (Triticum aestivum). In order to study the main physiological characterization, morphology and ultra-structure of chloroplast in a yellow-green leaf color mutant of common wheat, three near-isogenic lines (NILs) (a xantha line 1-20YY, a yellow-green line 1-20YG and a green line 1-20GG) which were derived from this yellow-green mutant were used by examination of the soluble substances content, a series of antioxidant enzymes activity, and morphology and ultra-structure of chloroplast. The results showed that the change of the leaf color had more influence on the content of soluble sugar and protein than that on the soluble proline. The content of soluble sugar and protein in the xantha line 1-20YY were significantly lower than in the green line 1-20GG and the yellow-green line 1-20YG except seedling stage (P＜0.05). And the content of soluble sugar and protein in the yellow-green line 1-20YG was no difference compared with the green line 1-20GG at all stages. Meanwhile, the yellowing of leaves also effected on activity of antioxidant enzymes. The influence on the activity of peroxidase (POD) was higher than that on the activity of superoxide dismutase (SOD) and hydrogen peroxidase (CAT). So this yellowing of leaves had serious influences to the content of soluble sugar and the activity of POD. From the chloroplast morphology, it could be found that the chloroplast shape of the xantha line 1-20YY was wad irregular and had a trend to the center of mesophyll cell. The chloroplast shape of the green line 1-20GG was nearly ellipse and the shape of chloroplast in 1-20YG line was almost spherical. The results of chloroplast ultra-structure with transmission electron microscopy revealed that the stroma thylakoids and grana thylakoids of the 1-20YY line were underdeveloped and their connection of each other were not tighter than that of 1-20YG line and 1-20GG line. There were no difference of the chloroplast ultra-structure between 1-20YG line and 1-20GG line. All these results provide a foundation for in-depth characterization of the formation mechanisms of the yellow-green leaf color mutant of wheat.
[1]安旭尧,刘聪,朱传杰,等. 2015.黄化小麦近等基因系遗传背景、光合和农艺性状分析.麦类作物学报[J].35(11):1476-1482.（An X Y, Liu C, Zhu C J, et al..2015. Evaluation and Analysis of the Main Photosynthetic Properties and Agronomic Characteristics of Three Xantha Wheat NILs[J]. Journal of Triticeae Crops.35(11):1476-1482.)
[2]曹莉,王辉,孙道杰,等.2008.一个新的小麦黄化突变体的遗传研究[J].遗传,30（12）:1603-1607.（Cao L, Wang H, Sun D J, et al.. 2008. Genetic analysis of a novel aurea mutant in wheat[J]. Hereditas, 30(12):1603-1607.
[3]曹莉,王辉,孙道杰,等.2010.小麦黄化突变体类囊体蛋白组分及叶绿素的合成特性[J].麦类作物学报.30(4):638-643.(Cao L, Wang H, Sun D J, et al.. 2010. Chloroplast Thylakoid Protein Composition and Characteristics of Chlorophyll Biosynthesis in a Novel Aurea Mutant of Wheat[J].Journal of Triticeae Crops,30(4):638-643. )
[4]曹莉,王辉,孙道杰,等.2006.小麦黄化突变体叶绿体超微结构研究[J].西北植物学报,26(11):2227-2230.(Cao L, Wang H, Sun D J. et al.. Chloroplast ultra-structure of a Xantha wheat mutant[J].Acta Bot Boreali-Occidentalla Sin, 26(11):2227-2230.)
[5]陈贵,康宗利,张立军.1998.低温胁迫对小麦生理生化特性的影响[J].麦类作物学报,18(3):42-43.(Chen G, Kang Z L, Zhang L J. 1998. Influence of physiological and biochemical characteristics of wheat under low temperature stress[J]. Tritical Crops,18(3):42-43.)
[6]姜春明,尹燕枰,刘霞,等.2007.不同耐热性小麦品种旗叶膜脂过氧化和保护酶活性对花后高温胁迫的响应[J].作物学报,33(1):143-148.(Jiang C M,Yin Y P,Liu X,et al.. 2007. Response of flag leaf lipid peroxidation and protective enzyme activity of wheat cultivars with different heat tolerance to high temperature stress after anthesis[J]. Acta Agron Sin, 3(1):143-148.)
[7]李合生.2000.植物生理生化实验原理和技术[M].北京:高等教育出版社,67-261(Li H S. 2000. Principle and Technology of Physiological and Biochemical Experiments of Plant[M]. BeiJing:Higher Education Press, 167-261)
[8]刘胜,魏祥进,邵高能,等.2013.一个水稻“斑马叶”叶色突变基因zebra leaf2(zl2)的图位克隆[J].中国水稻科学.27(3):231-239.（Liu S, Wei X J, Shao G N, et al.. 2013. Map-based Cloning of A ‘Zebra’ Leaf Mutant Gene zl2 in Rice[J]. Chin J Rice Sci, 27(3):231-239.)
[9]李美如,刘鸿先,王以柔.1996.细胞氧化应激机制与植物抗冷性机理的研究[J].生命科学,4:1-6.(Li M R, Liu H X, Wang Y R. 1996 .The Study on Oxidative Stress Mechanism of Cell and The Cold Resistance Mechanism[J]. Life Sciences, 4:1-6.)
[10]刘林,李明泽,王怀中,等.2010.不同发育时期板栗叶肉细胞中叶绿体淀粉和质体小球的变化[J].果树学报,27(1):131-134.(Liu L, Li M Z, Wang H Z, et al.. 2010. Changes of starch grains and plastoglobuli in choroplasts of mesophyll cells in Chinese chestnut leaf at different stages[J]. Journal of Fruit Science, 27(1):131-134.)
[11]谭炎宁,孙学武,袁定阳,等.2015.水稻单叶独立转绿型黄化突变体grc2的鉴定与基因精细定位[J].作物学报，41(6):831-837.(Tan Y N, Sun X W, Yuan D Y, et al.. 2015. Identification and fine mapping of Green-Revertible chlorina gene grc2 in rice(Oryza sativa L.)[J]. ACTA AGRONOMICA SINICA, 41(6):831-837.)
[12]王毅,杨宏福,李树德.1994.园艺植物冷害与抗冷性的研究[J].园艺学报,21(3)：239-244.(Wang Y, Yang H F, Li S D. 1994. The Research of Chilling Injury and Cold Rresistance of Plant[J].Acta Horticulturae Sinica, 21(3):239-244.)
[13]肖华贵,杨焕文,饶勇,等.2013甘蓝型油菜黄化突变体的叶绿体超微结构、气孔特征参数及光合特性[J].中国农业科学,46(4):715-727.(Xiao H G, Yang H W, Rao Y, et al.. 2013. Analysis of chloroplast ultrastructure,stonatal characteristic parameters and photosynthetic characteristics of chlorophyll-reduced mutant in Brassica napus L.[J]. Scientia Agricultura Sinica, 46(4):715-727.)
[14]杨冬业,张丽珍,徐淑庆.2012.西瓜不同生长阶段的过氧化氢酶活性研究[J].安徽农业科学,40(18):9604-9606.(Yang D Y, Zhang L Z, Xu S Q.2012. Study on Catalase Activity in Watermelon during Its Different Growth Periods[J]. Journal of Anhui Agricultural Sciences, 40(18):9604-9606.)
[15]于晶,张林,崔红,等.高寒地区冬小麦东农冬麦1号越冬前的生理生化特性[J].作物学报,2008,34(11):2019-2025.(Yu J, Zhang L, Cui H, et al.. 2008 Physiological and Biochemical Characteristics of Dongnongdongmai 1 before Wintering in High-Cold Area[J]. ACTA AGRONOMICA SINICA, 34(11):2019-2025.)
[16]Asakura,Y., S.Kikuchi and M.Nakai. 2008 . Non-identical contributions of two membrane-bound cpSRP components, cpFtsY and Alb3, to thylakoid biogenesis. Plant J, 56:1007-1017.
[17]Allen R D. 1995. Dissection of oxidative stress tolerance using transgentic plants[J]. Plant Physiol, 107:1049-1054.
[18]Back K H, Skinner D Z. 2006. Differential expression of manganese superoxide dismutase sequence variants in near isogentic lines of wheat during cold acclimation[J].Plant Cell Rep, 25:223-230. [19]Granick S, Gassman M L. 2000. Rapid regeneration of protochlorophyllide 650[J]. Plant Physiol, 45:201-205.
[20]Garaham D. Patterson B D. 1982. Responses of plants to low nonfreezing temperatures:proteins metabolism, and acctimation[J]. Annu Rev Plant Physiol, 33:347-372.
[21]Hui, Z., F.X.Tian, G.K.Wang and W.Wang. 2012. The antioxidantive defense system is involved in the delayed senescence in a wheat mutant tasg1[J]. Plant Cell Rep, 31:1073-1084.
[22]Jarvis, P., P.Dormann, C.A.Peto, J.Lutes, C.Benning and J.Chory. 2000. Galactolipid deficiency and abnormal chloroplast development in the Arabidopsis MGD synthase 1 mutant[J]. Proceedings of The National Academy of Sciences, 97(14):8175-8179.
[23]Kosuge, K., N.Watanabe and T.Kuboyama. 2011. Comparative genetic mapping of homoeologous genes for the chlorina phenotype in the genus Triticum[J]. Euphytica, 179:257-263.
[24]Liu, Z., S.W.Hong, M.Escobar, E.Vierling, D.L.Mitchell, D.W.Mount and J.D.Hall. 2003. Arabidopsis UVH6, a homolog of human XPD and yeast RAD3 DNA repair genes, functions in DNA repair and is essential for plant growth. Plant Physiol, 132(3):1405-1414.
[25]Li N, Jia J Z, Xia C,et al. 2013. Characterization and mapping of novel chlorophyll deficient mutant genes in durum wheat[J]. Breeding Science, 63(2):169-175.
[26]Luo Pei-Gao, Ren Zheng-Long. 2006. Wheat Leaf Chlorosis Controlled by a Single Recessive Gene[J]. Journal of Plant Physiology and Molecular Biology, 32(3):330-338.
[27]M.J.Ansari, A. Al-ghamdi, R. Kumar, et al. . 2013. Characterization and gene mapping of a chlorophyll-deficient mutant clm1 of Triticum monococcum L[J]. Biologia Plantarum. 57(3):442-448.
[28]Oster U, R Tanaka. 2000. Cloning and functional expression of the gene encoding the key enzyme for chlorophyll b biosynthesis (CAO) from Arabidopsis thaliana[J]. Plant Journal , 21(3):305-310.
[29]Schultes N P, R J H Sawers. 2000. Maize high chlorophyll fluorescent 60 mutation is caused by an Ac disruption of the gene encoding the chloroplast ribosomal small subunit protein 17[J]. Plant Journal,21(4):317-327.
[30]Zhang H T, Li J J, Yoo J H, et al. 2006. Rice Chlorina-1 and Chlorina-9 encode ChlD and ChlI subunits of Mg-chelatase, a key enzyme for chlorophyll synthesis and chloroplast development[J]. Plant Mol Biol, 62:325-337.
