陕糯1号与非糯小麦西农1330胚乳发育及淀粉形态、粒径分析

doi:10.3864/j.issn.0578-1752.2014.22.006

摘要/Abstract

摘要： 【目的】比较分析糯小麦陕糯1号和非糯小麦西农1330胚乳发育及淀粉形态、粒径的差异，为糯性与非糯性小麦的品质改良和开发利用提供理论依据。【方法】选用陕糯1号（糯性）和西农1330（非糯性）2个小麦品种，取发育过程中籽粒（花后5、8、12、15、18、21、25和28 d）液氮速冻后横断，经戊二醛和锇酸固定、磷酸缓冲液漂洗、丙酮梯度脱水、Epon812胶包埋后，采用超薄切片机切成1 μm厚的半薄切片，经1%甲苯胺蓝染色后采用光学显微镜观察胚乳细胞发育。提取2个小麦品种的淀粉粒，将其与成熟期籽粒横断面分别固定在样品台上，经离子溅射镀金后用扫描电镜观察成熟期胚乳结构及淀粉粒形态。采用MASTERSIZER-2000激光粒度仪分析淀粉粒粒径分布特征，每个样品重复3次，测定结果采用DPS统计软件进行数据统计分析，Sigmaplot 12.0作图。【结果】在小麦籽粒发育过程中，陕糯1号胚乳细胞体积大小及增大幅度均小于西农1330，发育早期，二者淀粉粒表面均能被甲苯胺蓝染色，发育中后期，陕糯1号淀粉粒仍能被染色，西农1330淀粉粒则不能被染色。与西农1330相比，陕糯1号胚乳细胞内蛋白基质较少，蛋白质与淀粉粒结合较疏松。陕糯1号B型淀粉粒形态多为不规则的多边形，西农1330B型淀粉粒大多呈圆球形，二者A型淀粉粒形态无明显差异。陕糯1号和西农1330淀粉粒粒径分布存在差异。陕糯1号淀粉粒体积分布呈四峰曲线变化，西农1330则呈双峰曲线变化，二者表面积分布均呈三峰曲线变化，数目分布均呈单峰曲线变化。陕糯1号A 型（>10 μm）淀粉粒体积、表面积百分比均低于西农1330，B型（<10 μm）淀粉粒体积、表面积百分比均高于西农1330，二者A、B型淀粉粒数目百分比无明显差异。陕糯1号中SB型（<1 μm）淀粉粒的体积、表面积、数目占总淀粉粒比例分别比西农1330低1.11%、11.60%和9.28%，LB型（1—10 μm）淀粉粒的体积、表面积、数目占总淀粉粒比例分别比西农1330高8.27%、15.88%、9.27%。陕糯1号具有少量LA（>53 μm）型淀粉粒，而西农1330没有。【结论】在小麦籽粒发育过程中，陕糯1号与西农1330的胚乳发育及淀粉形态存在明显差异。LB型淀粉粒对陕糯1号与西农1330 B型淀粉粒粒径分布影响较大。

关键词: 糯小麦, 胚乳, 淀粉, 粒径, 显微结构

Abstract: 【Objective】 The aim of this research is to study the endosperm development and morphological features of starch in waxy wheat Shannuo 1 and non-waxy wheat Xinong 1330, which will provide a theoretical basis for quality improvement of wheat breeding. 【Method】 Two wheat cultivars, Shannuo 1 (waxy wheat) and Xinong 1330 (non-waxy wheat) were used as materials in this paper. The development of endosperm cells were observed by optical microscope. In order to preserve the endosperm structure, the developing grains (5, 8, 12, 15, 18, 21, 25 and 28 d after fertilization) were immersed in glutaraldehyde and osmic acid after transected by freezing in liquid nitrogen. Before embedded with Epon812, samples were washed with phosphate buffer and dehydrated individually in a graded acetone solution. The semi-thin slices were obtained by Leica ULTRACUT slicer. Images were taken by optical microscope after stained by 1% toluidine. The cross sections of Shannuo 1 and Xinong 1330 grains and starch samples which isolated from the two wheat varieties were placed on an aluminum specimen holder, and sputter-coated with a thin ?lm of gold under vacuum condition. Samples were observed with a JEOL scanning electron microscope (JSM-6360LV, JEOL, Japan). Particle size distribution of the starch samples was measured by MASTERSIZER-2000 laser particle analyzer (Malvern UK company). All measurements were performed in triplicates. Analysis of variance (ANOVA) was performed using LSD’s test to compare treatment variations at a signi?cance level of P＜0.05 with DPS software. The data figures were performed by Sigmaplot 12.0 software. 【Result】 The results indicated that the endosperm cells of waxy wheat Shannuo 1 were smaller and developed slower than those of non-waxy wheat Xinong 1330 during grain development. In the early period of the endosperm development, the starch granules of Shannuo 1 and Xinong 1330 could be stained well by toluidine. However, in the late period of the grain development, the starch granules of Shannuo 1 could also be stained well, but Xinong 1330 can not. Compared with Xinong 1330, the endosperm of Shannuo 1 contained less protein matrix and the combination between protein and starch granules was much looser. The shape of B type starch granules of Shannuo 1 was irregular polygon, while that of Xinong 1330 was spherical. There was no significant difference in the shape of A type starch granules between Shannuo 1 and Xinong 1330. The size distribution of starch granules varied between waxy and non-waxy wheat. Volume distribution of Shannuo 1 starch granules showed a typical four-peak distribution, while that of Xinong 1330 showed a bimodal distribution. Granule surface area distribution of both cultivars indicated a typical three-peak distribution. The number distribution of granules in waxy and non-waxy starch was a typical unimodal distribution．The percentage of starch granules of Shannuo 1 and Xinong 1330 was nearly identical. In contrast, the percentage of the volume and the surface area of Shannuo 1 differed significantly from those of Xinong 1330. For A (>10 μm) type starch granules, the percentage of the volume and the surface area of Shannuo 1 were both lower than those of Xinong 1330. While for B (<10 μm) type starch granules, those of Shannuo 1 were higher than those of Xinong 1330. The volume, surface area and number of SB (<1 μm) type starch granules of Shannuo 1 were lower than those of Xinong 1330 by 1.11%, 11.60% and 9.28%, respectively, whereas the LB (1-10 μm) type starch granules of Shannuo 1 were higher than those of Xinong 1330 by 8.27%, 15.88% and 9.27%, respectively. A minority LA (>53 μm) type starch granules was found in Shannuo 1, but not in Xinong 1330. 【Conclusion】 In conclusion, the development of endosperm and the morphological features of starch granules of waxy wheat Shannuo 1 were significantly different from those of non-waxy wheat Xinong 1330 during grain development. The starch granules of LB type had considerable effects on the B type starch granules size distribution of Shannuo 1 and Xinong 1330.

Key words: waxy wheat, endosperm, starch, particle size, microstructure

余静，冉从福，李学军，邵慧，李立群. 陕糯1号与非糯小麦西农1330胚乳发育及淀粉形态、粒径分析[J]. 中国农业科学, 2014, 47(22): 4405-4416.

YU Jing, RAN Cong-fu, LI Xue-jun, SHAO Hui, LI Li-qun. Study on Endosperm Development and Morphological Features of Starch Granules in Waxy Wheat Shannuo 1 and Non-Waxy Wheat Xinong 1330[J]. Scientia Agricultura Sinica, 2014, 47(22): 4405-4416.

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参考文献

[1] 孟秀蓉, 熊飞, 孔妤, 陈永惠, 马守宝, 陆巍, 王忠. 强、中、弱筋小麦籽粒中淀粉、蛋白质积累和淀粉体发育的比较. 作物学报, 2009, 35(5): 962-966.

Meng X R, Xiong F, Kong Y, Chen Y H, Ma S B, Lu W, Wang Z. Comparison of starch, protein accumulation and amyloplast development in wheat cultivars with strong, medium and weak gluten. Acta Agronomica Sinica, 2009, 35(5): 962-966. (in Chinese)

[2] 戴忠民, 王振林, 张敏, 李文阳, 闫素辉, 蔡瑞国, 尹燕枰. 不同品质类型小麦籽粒淀粉粒度的分布特征. 作物学报, 2008, 34(3): 465-470.

Dai Z M, Wang Z L, Zhang M, Li W Y, Yan S H, Cai R G, Yin Y P. Starch granule size distribution in grains of strong and weak gluten wheat cultivars. Acta Agronomica Sinica, 2008, 34(3): 465-470. (in Chinese)

[3] Zeng J, Gao H, Li G L. Functional properties of wheat starch with different particle size distribution. Journal of the Science of Food and Agriculture, 2014, 94(1): 57-62.

[4] Kim H S, Huber K C. Physicochemical properties and amylopectin fine structures of A-and B-type granules of waxy and normal soft wheat starch. Journal of Cereal Science, 2010, 51(3): 256-264.

[5] Li W H, Shan Y L, Xiao X L, Luo Q G, Zheng J M, Ouyuan S H, Zhang G Q. Effect of nitrogen and sulfur fertilization on accumulation characteristics and physicochemical properties of A- and B-wheat starch. Journal of Agricultural and Food Chemistry, 2013, 61(10): 2418-2425.

[6] 李继刚, 梁荣奇, 张义荣, 李保云, 刘广田. 糯性普通小麦的产生及其淀粉特性研究. 麦类作物学报, 2001, 21(2): 10-13.

Li J G, Liang R Q, Zhang Y R, Li B Y, Liu G T. Production of waxy common wheat and its starch properties. Journal of Triticeae Crops, 2001, 21(2): 10-13. (in Chinese)

[7] 张晓, 高德荣, 吕国峰, 吴宏亚, 张伯桥, 李曼. 糯小麦与其它作物淀粉特性的比较研究. 中国农业科学, 2013, 46(11): 2183-2190.

Zhang X, Gao D R, Lü G F, Wu H Y, Zhang B Q, Li M. Comparison of the starches properties of waxy wheat and other crops. Scientia Agricultura Sinica, 2013, 46(11): 2183-2190. (in Chinese)

[8] Bhattacharya M, Erazo-Castrejón S V, Doehlert D C, McMullen M S. Staling of bread as affected by waxy wheat flour blends. Cereal Chemistry, 2002, 79(2): 178-182.

[9] Sajilata M G., Singhal R S, Kulkarni P R. Resistant starch–a review. Comprehensive Reviews in Food Science and Food Safety, 2006, 5(1): 1-17.

[10] 刘智, 王玲玲, 张二金, 翟干干, 熊飞, 张琛, 王忠. 不同类型专用小麦品种胚乳发育的比较研究. 麦类作物学报, 2011, 31(1): 70-76.

Liu Z, Wang L L, Zhang E J, Zhai G G, Xiong F, Zhang C, Wang Z. Study on endosperm development between wheat cultivars for different uses. Journal of Triticeae Crops, 2011, 31(1): 70-76. (in Chinese)

[11] 陈义芳, 张静, 周卫东, 张彪, 马雷, 韦存虚. 不同品质类型小麦籽粒结构的观察比较. 电子显微学报, 2006, 25(3): 271-274.

Chen Y F, Zhang J, Zhou W D, Zhang B, Ma L, Wei C X. Structural differences in the mature endosperms of wheat with different quality. Journal of Chinese Electron Microscopy Society, 2006, 25(3): 271-274. (in Chinese)

[12] 袁翠平, 田纪春. 小麦籽粒硬度与胚乳显微结构关系研究. 中国粮油学报, 2004, 19(2): 28-31.

Yuan C P, Tian J C. Study on relationship between hardness of wheat grain and microstructure of endosperm. Journal of the Chinese Cereals and Oils Association,2004, 19(2): 28-31. (in Chinese)

[13] Peterson D G, Fulcher R G. Variation in minnesota HRS wheats: Starch granule size distribution. Food Research International, 2001, 34(4): 357-363.

[14] Bechtel D B, Zayas I, Kaleikau L, Pommeranz Y. Size distribution of wheat starch granules during endosperm development. Cereal Chemistry, 1990, 67: 59-63.

[15] Yoo S H, Jane J. Structural and physical characteristics of waxy and other wheat starches. Carbohydrate Polymers, 2002, 49: 297-305.

[16] 蔡瑞国, 张敏, 朱桓, 武宝悦, 李彦生, 王振林. 糯小麦籽粒淀粉粒度分布特征. 麦类作物学报, 2010, 30(2): 254-258.

Cai R G, Zhang M, Zhu H, Wu B Y, Li Y S, Wang Z L. Starch granule size distribution in grains of waxy wheat. Journal of Triticeae Crops, 2010, 30(2): 254-258. (in Chinese)

[17] Zhang H, Zhang W, Xu C Z, Zhou X. Morphological features and physicochemical properties of waxy wheat starch. International Journal of Biological Macromolecules, 2013, 62: 304-309.

[18] Hansen L E, Jackson D S, Wehling R L, Wilson J D, Graybosch R A. Functionality of native tetraploid wheat starches: Effects of waxy loci alleles and amylose concentration in blends. Journal of Cereal Science, 2010, 52(1): 39-45.

[19] 熊飞, 王忠, 陈刚, 王珏, 李波. 不同专用小麦胚乳细胞淀粉体的比较研究. 扬州大学学报: 农业与生命科学版, 2005, 26(1): 73-75.

Xiong F, Wang Z, Chen G, Wang Y, Li B. A comparison study on the amyloplasts of endosperm cells in the different and special wheat. Journal of Yangzhou University: Agricultural and Life Science Edition, 2005, 26(1): 73-75. (in Chinese)

[20] 周竹青, 朱旭彤, 王维金, 兰盛银. 不同粒型小麦品种胚乳淀粉体的扫描电镜观察. 电子显微学报, 2001, 20(3): 178-184.

Zhou Z Q, Zhu X T, Wang W J, Lan S Y. Observation on the amyloplasts in endosperm of wheat varieties with different kernel types by scanning electron microscope. Journal of Chinese Electron Microscopy Society, 2001, 20(3): 178-184. (in Chinese)

[21] Ma H, Zhang X, Wang C G, Gao D R, Zhang B Q, Lv G F, Wu R L, Cheng X M, Wang X, Cheng S H, Bie T D. Effect of Wx genes on amylose content, physicochemical properties of wheat starch, and the suitability of waxy genotype for producing Chinese crisp sticks. Journal of Cereal Science, 2013, 58(1): 140-147.

[22] 张晶. 陕西小麦糯蛋白和硬度蛋白的基因组成分析[D]. 西安: 西北农林科技大学, 2011.

Zhang J. Identification of genotypes for waxy protein and puroindoline protein in Shaanxi wheat[D], Xi’an: Northwest Agricultural and Forest University, 2011. (in Chinese)

[23] Peng M S, Gao M, Abdel-Aal E S M. Separation and characterization of A-and B-type starch granules in wheat endosperm. Cereal Chemistry, 1999, 76: 375-379.

[24] 盛婧, 郭文善, 朱新开, 封超年, 彭永欣. 不同专用类型小麦籽粒淀粉粒形成过程. 作物学报, 2004, 30(9): 953-954.

Sheng J, Guo W S, Zhu X K, Feng C N, Peng Y X. Starch granules development in grain endosperm of wheat for different end uses. Acta Agronomica Sinica, 2004, 30(9): 953-954. (in Chinese)

[25] 王玲玲, 刘智, 熊飞, 李栋梁, 周卫东, 陈义芳, 王忠. 固定和染色方法对小麦胚乳细胞结构显示的影响. 作物学报, 2012, 38(9): 1688-1697.

Wang L L, Liu Z, Xiong F, Li D L, Zhou W D, Chen Y F, Wang Z. Effect of fixation and staining methods on structure observation of endosperm cell of wheat. Acta Agronomica Sinica, 2012, 38(9): 1688-1697. (in Chinese)

[26] 李春燕, 封超年, 王亚雷, 张容, 郭文善, 朱新开, 彭永欣. 不同小麦品种支链淀粉链长分配及其与淀粉理化特性的关系. 作物学报, 2007, 33(8): 1240-1245.

Li C Y, Feng C N, Wang Y L, Zhang R, Guo W S, Zhu X K, Peng Y X. Chain length distribution of debauched amylopectin and its relationship with physicochemical properties of starch in different wheat cultivar. Acta Agronomica Sinica, 2007, 33(8): 1240-1245. (in Chinese)

[27] Jane J, Chen Y Y, Lee L F, Mcpherson A E, Wong K S, Radosavljevic M, Kasemsuwan T. Effects of amylopectin branch chain length and amylase content on the gelatinization and pasting properties of starch. Cereal Chemistry, 1999, 76(5): 629-637.

[28] Svihus B, Uhlen A K, Harstad O M. Effect of starch granule structure, associated components and processing on nutritive value of cereal starch: A review. Animal Feed Science and Technology, 2005, 122(3): 303-320.

[29] Baldwin P M. Starch granule-associated proteins and polypeptides: A review. Starch, 2001, 53: 475-503.

[30] 曹颖妮, 胡卫国, 王根平, 刘录祥, 王成社. 糯性和非糯性小麦灌浆期胚乳直/支链淀粉积累及其相关酶活性研究. 西北植物学报, 2010, 30(10): 1995-2001.

Cao Y N, Hu W G, Wang G P, Liu L X, Wang C S. Dynamic changes of starch accumulation and enzymes relating to starch biosynthesis of kernel during grain filling in waxy and non-waxy winter wheat. Acta Botanica Boreali- Occidentalia Sinica, 2010, 30(10): 1995-2001. (in Chinese)

[31] Cao Y N, Hu W G, Wang C S. Relationship among the key enzymatic activities involved in starch synthesis and amylopectin chain distributions in developing wheat grain. African Journal of Biotechnology, 2012, 11(4): 805-814.

[32] 闫素辉. 小麦胚乳淀粉合成、粒度分布特征及对花后高温的响应[D]. 泰安: 山东农业大学, 2009.

Yan S H. Starch synthesis distribution of developing wheat endosperm in response to postanthesis high temperature[D]. Taian: Dissertation of Shandong Agricultural University, 2009. (in Chinese)

[33] Li W H, Shan Y L, Xiao X L, Luo Q G, Zheng J M, Ouyang S H, Zhang G Q. Physicochemical properties of A-and B-starch granules isolated from hard red and soft red winter wheat. Journal Agriculture Food Chemistry, 2013, 61: 6477-6484.