Transcriptome Analysis of Sugarcane Young Leaves and Protoplasts after Enzymatic Digestion.
Transcriptome Analysis of Sugarcane Young Leaves and Protoplasts after Enzymatic Digestion.
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酶消化后甘蔗幼叶和原生质体的转录组分析
DOI：
10.3390/life12081210
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发表时间：
2022-08-09
期刊：
影响因子：
3.2
通讯作者：
Li, Suli
中科院分区：
文献类型：
--
作者：
Zhang, Demei;Wang, Rui;Han, Shijian;Li, Zhigang;Xiao, Jiming;Li, Yangrui;Wang, Lingqiang;Li, Suli
甘蔗体细胞杂交可以突破传统育种中远亲种之间遗传不亲和的障碍。然而，甘蔗原生质体再生的分子机制和原生质体制备的条件仍不清楚。本研究以甘蔗新台糖22号（ROC 22）幼叶为材料，采用2%纤维素酶+0.5%果胶酶+0.1%游离酶+0.3%半纤维素酶（pH = 5.8）进行酶解，原生质体活力达90%以上。对酶切后的甘蔗幼叶和原生质体进行转录组测序，分析酶切前后体细胞基因表达的差异。共获得117，411条unigenes和43，460条差异表达基因，其中21，123条上调，22，337条下调。43，460个差异表达基因（DEG）的GO术语分为三大类：生物学过程、细胞组分和分子功能，涉及发育过程、生长、细胞增殖、转录调节活性、信号转导活性、抗氧化活性、氧化胁迫、激酶活性、细胞周期、细胞分化、植物激素信号转导、甘蔗幼叶经酶消化后，GAUT、CESA、PSK、CyclinB、CyclinA、CyclinD 3和cdc 2等与植株再生相关的基因的表达量分别显著下调至幼叶的65%、47%、2%、18.60%、21.32%、52%和45%。酶解后，Aux/IAA表达量较幼叶上调，Aux/IAA表达量是幼叶的3.53倍。与幼叶相比，酶解后这些关键基因发生了显著变化。这些结果表明，体细胞酶消化过程可能在一定程度上影响杂合细胞的再生。
Sugarcane somatic cell hybridization can break through the barrier of genetic incompatibility between distantly related species in traditional breeding. However, the molecular mechanisms of sugarcane protoplast regeneration and the conditions for protoplast preparation remain largely unknown. In this study, young sugarcane (ROC22) leaves were enzymatically digested, and the viability of protoplasts reached more than 90% after enzymatic digestion (Enzymatic combination: 2% cellulase + 0.5% pectinase + 0.1% dissociative enzyme + 0.3% hemicellulase, pH = 5.8). Transcriptome sequencing was performed on young sugarcane leaves and protoplasts after enzymatic digestion to analyze the differences in gene expression in somatic cells before and after enzymatic digestion. A total of 117,411 unigenes and 43,460 differentially expressed genes were obtained, of which 21,123 were up-regulated and 22,337 down-regulated. The GO terms for the 43,460 differentially expressed genes (DEGs) were classified into three main categories: biological process, cellular component and molecular function, which related to developmental process, growth, cell proliferation, transcription regulator activity, signal transducer activity, antioxidant activity, oxidative stress, kinase activity, cell cycle, cell differentiation, plant hormone signal transduction, and so on. After enzymatic digestion of young sugarcane leaves, the expressions of GAUT, CESA, PSK, CyclinB, CyclinA, CyclinD3 and cdc2 genes associated with plant regeneration were significantly down-regulated to 65%, 47%, 2%, 18.60%, 21.32%, 52% and 45% of young leaves, respectively. After enzymatic digestion, Aux/IAA expression was up-regulated compared with young leaves, and Aux/IAA expression was 3.53 times higher than that of young leaves. Compared with young leaves, these key genes were significantly changed after enzymatic digestion. These results indicate that the process of somatic enzymatic digestion process may affect the regeneration of heterozygous cells to a certain extent.
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影响因子：
27.5
作者：
Caffall, Kerry H.;Pattathil, Sivakumar;Mohnen, Debra
通讯作者：
Mohnen, Debra
影响因子：
6.5
作者：
Eun, CH;Ko, SM;Kamada, H
通讯作者：
Kamada, H
DOI：
10.1073/pnas.94.24.13357
发表时间：
1997-11-25
影响因子：
11.1
作者：
Matsubayashi, Y;Takagi, L;Sakagami, Y
通讯作者：
Sakagami, Y
影响因子：
2.5
作者：
Negis, Yesim;Unal, Aysegul Yildiz;Karabay, Arzu
通讯作者：
Karabay, Arzu
影响因子：
11.4
作者：
De Veylder, L;Beeckman, T;Inzé, D
通讯作者：
Inzé, D