Development and Application of an Efficient Method for the Amylose/ Amylopectin Ratio Determination in Potato Tubers

doi:10.16420/j.issn.0513-353x.2020-0446

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (3): 600-608.doi: 10.16420/j.issn.0513-353x.2020-0446

• New Technology and New Mathod • Previous Articles Next Articles

Development and Application of an Efficient Method for the Amylose/ Amylopectin Ratio Determination in Potato Tubers

SONG Yuhao, SHI Weiling, ZHANG Jiao, DING Zhenyu, PENG Hongxian, JIANG Chunyan, MA Qiuqin, LI Zhijing, ZHAO Yong, TANG Daobin, ZHANG Kai, WANG Jichun, LIU Xun()

Chongqing Key Laboratory of Biology and Genetic Improvement in Tuber and Root Crops,College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China

Received:2020-07-10 Online:2021-03-25 Published:2021-04-02
Contact: LIU Xun E-mail:liuxun828@swu.edu.cn

Abstract

Abstract:

Potato starch composition（amylose/amylopectin,AM/AP）influences functional characteristics for food and other industrial applications. The determination of AM/AP ratio is of great significance for rational utilization of starch and breeding special varieties with specific starch components. The dual-wavelength method for determination of AM and AP contents was established in a 96-well plate format firstly. It was identified that the AM contents was dramatically overestimated when the AM/AP ratio of tested samples exceeded the AM/AP ratio（33.3%）of the standard curve. Furthermore,the new standard curve of potato AM/AP ratio and wavelength under the maximum absorption peak were established:y= 601.88x^0.0215,R²= 0.9999. This method further improves the accuracy and is applicable over large ranges of AM/AP ratio needed for screening. This method was used to determine the AM/AP ratio from 198 potato varieties（lines）and a few transgenic tubers with high amylose contents. The result revealed that the range of AM contents in our potato genetic resources is 17.4%-33.3%. In conclusion,a simple and efficient method for the accurate estimation of the potato starch composition was established which is especially suitable for the detection of genetic breeding materials and improvement starch composition lines.

Key words: potato, starch component, detection system, amylose, amylopectin

CLC Number:

S532

SONG Yuhao, SHI Weiling, ZHANG Jiao, DING Zhenyu, PENG Hongxian, JIANG Chunyan, MA Qiuqin, LI Zhijing, ZHAO Yong, TANG Daobin, ZHANG Kai, WANG Jichun, LIU Xun. Development and Application of an Efficient Method for the Amylose/ Amylopectin Ratio Determination in Potato Tubers[J]. Acta Horticulturae Sinica, 2021, 48(3): 600-608.

Figures/Tables 7

References 26

[1]	Copeland L, Blazek J, Salman H, Tang M C. 2009. Form and functionality of starch. Food Hydrocolloids, 23 (6):1527-1534. doi: 10.1016/j.foodhyd.2008.09.016 URL
[2]	Dai Shuang, Cheng Dun-gong, Li Hao-sheng, Liu Ai-feng, Liu Jian-jun, Zhao Zhen-dong, Song Jian-min. 2008. Simultaneous and rapid spectrophotometric determination of amylose,amylopectin,and total starch in wheat grain. Journal of Triticeae Crops, 28 (3):442-447. (in Chinese)
	戴双, 程敦公, 李豪圣, 刘爱峰, 刘建军, 赵振东, 宋健民. 2008. 小麦直、支链淀粉和总淀粉含量的比色快速测定研究. 麦类作物学报, 28 (3):442-447.
[3]	Delwiche S R, Bean M M, Miller R E, Webb B D, Williams P C. 1995. Apparent amylose content of milled rice by near-infrared reflectance spectroscopy. Cereal Chemistry, 72 (2):182-187.
[4]	Fajardo D, Jayanty S S, Jansky S H. 2013. Rapid high throughput amylose determination in freeze dried potato tuber samples. Journal of Visualized Experiments, 80:e50407-e504407.
[5]	Fei Y Y, Yang J, Wang F Q, Fan F J, Li W Q, Wang J, Xu Y, Zhu J Y, Zhong W G. 2019. Production of two elite glutinous rice varieties by editing Wx gene. Rice Science, 26 (2):118-124. doi: 10.1016/j.rsci.2018.04.007 URL
[6]	Guan S Y, Wang P W, Liu H J, Liu G N, Ma Y Y, Zhao L N. 2011. Production of high-amylose maize lines using RNA interference in sbe2a. African Journal of Biotechnology, 10 (68):15229-15237.
[7]	Guo Zhi-hong, Zhang Jin-wen, Wang Di, Chen Zheng-hua. 2008. Using RNAi technology to produce high-amylose potato plants. Scientia Agricultura Sinica, 41 (2):494-501. (in Chinese)
	郭志鸿, 张金文, 王蒂, 陈正华. 2008. 用RNA干扰技术创造高直链淀粉马铃薯材料. 中国农业科学, 41 (2):494-501.
[8]	Hofvander P, Andersson M, Larsson C, Larsson H. 2004. Field performance and starch characteristics of high-amylose potatoes obtained by antisense gene targeting of two branching enzymes. Plant Biotechnology Journal, 2 (4):311-320. pmid: 17134392
[9]	Hovenkamp-Hermelink J, Devries J, Adamse P, Jacobsen E, Witholt B, Feenstra W. 1988. Rapid estimation of the amylose amylopectin ratio in small amounts of tuber and leaf tissue of the potato. Potato Research, 31:241-246. doi: 10.1007/BF02365532 URL
[10]	Kaufman R C, Wilson J D, Bean S R, Herald T J, Shi Y C. 2015. Development of a 96-well plate iodine binding assay for amylose content determination. Carbohydrate Polymers, 115:444-447. doi: 10.1016/j.carbpol.2014.09.015 pmid: 25439917
[11]	Kitahara K, Hamasuna K, Nozuma K, Otani M, Hamada T, Shimada T, Fujita K, Suganuma T. 2007. Physicochemical properties of amylose-free and high-amylose starches from transgenic sweetpotatoes modified by RNA interference. Carbohydrate Polymers, 69 (2):233-240. doi: 10.1016/j.carbpol.2006.09.025 URL
[12]	Luo Fa-xing, Huang Qiang, Zhang Le-xing. 2007. The study and development of waxy potato. Food Research and Development, 28 (7):148-149. (in Chinese)
	罗发兴, 黄强, 张乐兴. 2007. 蜡质马铃薯淀粉的研究开发. 食品研究与开发, 28 (7):148-149.
[13]	Lü Wen-he, Huo Dan-dan, Li Yong, Lü Dian-qiu, Bai Ya-mei, Su Fei-fei. 2017. Comparison of starch contents and starch synthase activities in different parts of potato tuber. Journal of Northeast Agricultural University, 48 (6):1-8. (in Chinese)
	吕文河, 霍丹丹, 李勇, 吕典秋, 白雅梅, 宿飞飞. 2017. 马铃薯块茎不同部位淀粉含量及淀粉合成关键酶活性差异比较. 东北农业大学学报, 48 (6):1-8.
[14]	Ngobese N Z, Workneh T S, Alimi B A, Tesfay S. 2017. Nutrient composition and starch characteristics of eight European potato cultivars cultivated in South Africa. Journal of Food Composition and Analysis, 55:1-11. doi: 10.1016/j.jfca.2016.11.002 URL
[15]	Schirmer M, Hochstotter A, Jekle M, Arendt E, Becker T. 2013. Physicochemical and morphological characterization of different starches with variable amylose/amylopectin ratio. Food Hydrocolloids, 32 (1):52-63. doi: 10.1016/j.foodhyd.2012.11.032 URL
[16]	Schwall G P, Safford R, Westcott R J, Jeffcoat R, Tayal A, Shi Y C, Gidley M J, Jobling S A. 2000. Production of very-high-amylose potato starch by inhibition of SBE A and B. Nature Biotechnology, 18 (5):551-554. doi: 10.1038/75427 URL
[17]	Šimkováa D, Lachman J, Hamouz K, Vokál B. 2013. Effect of cultivar,location and year on total starch,amylose,phosphorus content and starch grain size of high starch potato cultivars for food and industrial processing. Food Chemistry, 141 (4):3872-3880. doi: 10.1016/j.foodchem.2013.06.080 pmid: 23993560
[18]	Singh J, Kaur L. 2016. Advances in potato chemistry and technology. 2nd ed. Amsterdam: Elsevier Press.
[19]	Su Fei-fei, Shi Ying, Liang Jing, Zhao Gui-yun, Chen Yi-li. 2006. Evaluation of different potato varieties for starch content,starch yield and starch component. Chinese Potato Journal, 20 (1):16-18. (in Chinese)
	宿飞飞, 石瑛, 梁晶, 赵桂云, 陈伊里. 2006. 不同马铃薯品种淀粉含量、淀粉产量及淀粉组成的评价. 中国马铃薯, 20 (1):16-18.
[20]	Sun Y W, Jiao G A, Liu Z P, Zhang X, Li J Y, Guo X P, Du W M, Du J L, Francis F, Zhao Y D, Xia L Q. 2017. Generation of high-amylose rice through CRISPR/Cas9-mediated targeted mutagenesis of starch branching enzymes. Frontiers in Plant Science, 8:298.
[21]	Willims P C, Kuzina F D, Hlynka I. 1970. A rapid colorimetrie pr ocedure for estimating the amylose content of starches and flours. Cereal Chemistry, 47:411-420.
[22]	Yang Huan-chun, Li Yong, Lü Wen-he, Du Ying-qiu, Shan Hong, Kong Bao-hua. 2015. Comparison of determination method for amylose content in potato tubers. Chinese Potato Journal, 146 (6):37-41. (in Chinese)
	杨焕春, 李勇, 吕文河, 杜英秋, 单宏, 郭保华. 2015. 马铃薯块茎直链淀粉含量检测方法的比较. 中国马铃薯, 146 (6):37-41.
[23]	Zeng Fan-kui, Zhao Xin, Zhou Tian-hong, Liu Gang. 2012. Dual-wavelength colorimetric method for measuring amylose and amylopectin contents of potato starch. Modern Food Science and Technology, 28 (1):119-122. (in Chinese)
	曾凡逵, 赵鑫, 周添红, 刘刚. 2012. 双波长比色法测定马铃薯直链/支链淀粉含量. 现代食品科技, 28 (1):119-122.
[24]	Zhang Jiao. 2018. Screening of potato breeding parents of stable food and establishment of shade tolerance evaluation system[M. D. Dissertation]. Chongqing:Southwest University. (in Chinese)
	张娇. 2018. 主食化马铃薯育种亲本的筛选及耐荫性评价体系的建立[硕士论文]. 重庆:西南大学.
[25]	Zhao Y J, Li N, Li B, Li Z X, Xie G N, Zhang J. 2015. Reduced expression of starch branching enzyme IIa and IIb in maize endosperm by RNAi constructs greatly increases the amylose content in kernel with nearly normal morphology. Planta, 241 (2):449-461. doi: 10.1007/s00425-014-2192-1 URL
[26]	Zhu T, Jackson D S, Wehling R L, Geera B. 2008. Comparison of amylose determination methods and the development of a dual wavelength iodine binding technique. Cereal Chemistry, 85:51-58. doi: 10.1094/CCHEM-85-1-0051 URL

AM + AP标准样品/μL Standard AM + AP		测定值/（μg·mL^-1） Measured value					回收率/% Recycling rate
AM + AP标准样品/μL Standard AM + AP		AM		AP	TS		AM		AP		TS
5.0 + 95.0 10.0 + 90.0 20.0 + 80.0 30.0 + 70.0 40.0 + 60.0 50.0 + 50.0 60.0 + 40.0 70.0 + 30.0 80.0 + 20.0 90.0 + 10.0		5.2 ± 0.1 10.2 ± 0.1 20.9 ± 0.2 35.5 ± 0.4 51.7 ± 0.2 63.2 ± 0.1 71.6 ± 0.2 84.9 ± 0.4 93.3 ± 0.3 98.8 ± 0.4		93.2 ± 0.2 86.4 ± 0.5 80.1 ± 0.2 71.5 ± 0.2 61.4 ± 0.2 51.7 ± 0.1 41.1 ± 0.1 34.2 ± 0.1 24.5 ± 0.2 21.0 ± 0.5	98.4 ± 0.2 96.6 ± 0.4 101.0 ± 0.3 107.0 ± 0.1 113.1 ± 0.2 114.9 ± 0.2 112.7 ± 0.2 119.1 ± 0.3 117.8 ± 0.2 119.8 ± 0.4		104.1 102.1 104.5 118.2 129.3 126.4 119.3 121.3 116.6 109.8		98.1 95.9 100.1 102.2 102.3 103.4 102.8 114.0 122.5 210.0		98.8 96.6 101.0 107.0 113.1 114.9 112.7 119.1 117.8 119.8

AM + AP标准样品/μL Standard AM + AP		测定值/（μg·mL^-1） Measured value					回收率/% Recycling rate
AM + AP标准样品/μL Standard AM + AP		AM		AP	TS		AM		AP		TS
5.0 + 95.0 10.0 + 90.0 20.0 + 80.0 30.0 + 70.0 40.0 + 60.0 50.0 + 50.0 60.0 + 40.0 70.0 + 30.0 80.0 + 20.0 90.0 + 10.0		5.2 ± 0.1 10.2 ± 0.1 20.9 ± 0.2 35.5 ± 0.4 51.7 ± 0.2 63.2 ± 0.1 71.6 ± 0.2 84.9 ± 0.4 93.3 ± 0.3 98.8 ± 0.4		93.2 ± 0.2 86.4 ± 0.5 80.1 ± 0.2 71.5 ± 0.2 61.4 ± 0.2 51.7 ± 0.1 41.1 ± 0.1 34.2 ± 0.1 24.5 ± 0.2 21.0 ± 0.5	98.4 ± 0.2 96.6 ± 0.4 101.0 ± 0.3 107.0 ± 0.1 113.1 ± 0.2 114.9 ± 0.2 112.7 ± 0.2 119.1 ± 0.3 117.8 ± 0.2 119.8 ± 0.4		104.1 102.1 104.5 118.2 129.3 126.4 119.3 121.3 116.6 109.8		98.1 95.9 100.1 102.2 102.3 103.4 102.8 114.0 122.5 210.0		98.8 96.6 101.0 107.0 113.1 114.9 112.7 119.1 117.8 119.8

Development and Application of an Efficient Method for the Amylose/ Amylopectin Ratio Determination in Potato Tubers

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