Chromoplast Isolation and Its Proteomic Analysis from Cassava Storage Roots

DENG Chang-Zhe1,2 YAO Hui1 AN Fei-Fei1 LI Kai-Mian1 CHEN Song-Bi1

(1.Tropical Crops Genetic resources Institute, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Ministry of Agriculture for Germplasm Resources Conservation and Utilization of Cassava, Danzhou, China 571737)
(2.Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, China 570228)

【Abstract】Chromoplasts are the sites to store carotenoids and regulate a variety of physiological and biochemical processes in the storage roots of cassava (Manihot esculenta Crantz). In the present study, it was found that Percoll density gradient centrifugation was suitable for isolating the chromoplasts from cassava storage roots. Rich and intact chromoplasts were located in 40% to 50% layer of Percoll, the expression level of Vdac1, a mitochondrial marker, was the lowest and the expression level of RbcL, a plastid marker, was the highest compared with those in other layers using Western blot. Thirty-four differentially expressed proteins were detected in SC9, in which 17 were up-regulated, and the others were down-regulated. The differential proteins related to carbohydrate and energy metabolism accounted for the highest proportion. The STRING protein-protein interaction network showed that Enolase 2 and Elongation Factor were the hub proteins, which played the key roles in the whole regulatory network. Quantitative analysis by qRT-PCR confirmed the Enolase 2 expression was more significantly up-regulated in the high carotenoid cassava cultivar than in the low carotenoid cassava cultivar SC6068. These two proteins may be the key points for affecting the carotenoid content between SC9 and SC6068.

【Keywords】 Cassava storage root; Chromoplast protein; Isolation method; Proteomics;


【Funds】 National Natural Science Foundation of China (31271776) supported by the National Natural Science Foundation of China (31271776) Initial Fund of High-level Creative Talents in Hainan Province (2012–2016) the Initial Fund of High-level Creative Talents in Hainan Province (2012–2016)

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This Article


CN: 11-1809/S

Vol 43, No. 09, Pages 1290-1299

September 2017


Article Outline


  • 1 Materials and methods
  • 2 Results and analyses
  • 3 Discussion
  • 4 Conclusions
  • References