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The seed oil of  Jatropha curcas  is considered a potential bioenergy source that could replace fossil fuels. However, the seed yield of  Jatropha  is low and has yet to be improved. We previously reported that exogenous cytokinin treatment increased the seed yield of  Jatropha . Cytokinin levels are directly regulated by isopentenyl transferase (IPT), cytochrome P450 monooxygenase, family 735, subfamily A (CYP735A), and cytokinin oxidase/dehydrogenase (CKX). In this study, we cloned six  IPT  genes, one  JcCYP735A  gene, and seven  JcCKX  genes. The expression patterns of these 14 genes in various organs were determined using real-time quantitative PCR.  JcIPT1  was primarily expressed in roots and seeds,  JcIPT2  was expressed in roots, apical meristems, and mature leaves,  JcIPT3  was expressed in stems and mature leaves,  JcIPT5  was expressed in roots and mature leaves,  JcIPT6  was expressed in seeds at 10 days after pollination, and  JcIPT9  was expressed in mature leaves.  JcCYP735A  was mainly expressed in roots, flower buds, and seeds. The seven  JcCKX  genes also showed different expression patterns in different organs of  Jatropha . In addition, CK levels were detected in flower buds and seeds at different stages of development. The concentration of N 6 -(Δ 2 -isopentenyl)-adenine (iP), iP-riboside, and  trans -zeatin (tZ) increased with flower development, and the concentration of iP decreased with seed development, while that of tZ increased. We further analyzed the function of  JcCYP735A  using the CRISPR-Cas9 system, and found that the concentrations of tZ and tZ-riboside decreased significantly in the  Jccyp735a  mutants, which showed severely retarded growth. These findings will be helpful for further studies of the functions of cytokinin metabolic genes and understanding the roles of cytokinins in  Jatropha  growth and development.

peerj

Identification and expression analysis of cytokinin metabolic genes IPTs, CYP735A and CKXs in the biofuel plant Jatropha curcas