Knockdown of a cellulose synthase gene BoiCesA affects the leaf anatomy, cellulose content and salt tolerance in broccoli.
Cellulose is the major component of cell wall materials. A 300 bp specific fragment from the cDNA fragment was chosen to insert into vector pFGC1008 at forward and reverse orientations to construct the recombinant RNAi vector. Knockdown of BoiCesA caused "dwarf" phenotype with smaller leaves and a loss of the content of cellulose. Moreover, RT-PCR analysis confirmed that the expression of the RNAi apparatus could repress expression of the CesA gene. Meanwhile, examination of the leaves from the T3 of RNAi transformants indicated reduction of cell expansion in vascular bundles, particularly on their abaxial surface. The proline and soluble sugar content increased contrarily. Under the salt stress, the T3 of RNAi plants showed significant higher resistance. The expression levels of some salt tolerance related genes (BoiProH, BoiPIP2;2, BoiPIP2;3) were significantly changed in T3 of RNAi plants. The results showed that the hairpin structure of CesA specific fragment inhibited the endogenous gene expression and it was proved that the cDNA fragment was relevant to the cellulose biosynthesis. Moreover, modulation cellulose synthesis probably was an important influencing factor in polysaccharide metabolism and adaptations of plants to stresses. This will provide technological possibilities for the further study of modulation of the cellulose content of crops.
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Shuangtao
S
Li
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Lei
L
Zhang
College of Horticulture, China Agricultural University, 100193, Beijing, China.|Horticulture Research Institute, Shanghai Academy Agricultural Sciences, Shanghai 201403, China.
Ying
Y
Wang
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Fengfeng
F
Xu
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Mengyun
M
Liu
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Peng
P
Lin
School of Agriculture, Virginia State University, PO Box 9061, Petersburg, VA23806, USA.
Shuxin
S
Ren
Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, Changchun 130033, China.
Rui
R
Ma
College of Horticulture, China Agricultural University, 100193, Beijing, China.
Yang-Dong
YD
Guo
D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics*; D000595:Amino Acid Sequence; D001937:Brassica / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D002473:Cell Wall / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D055785:Gene Knockdown Techniques; D017343:Genes, Plant; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics; D010368:Pectins / Q000378:metabolism; D010641:Phenotype; D018515:Plant Leaves / Q000033:anatomy & histology* / Q000187:drug effects / Q000648:ultrastructure; D030821:Plants, Genetically Modified; D011392:Proline / Q000378:metabolism; D034622:RNA Interference; D017422:Sequence Analysis, DNA; D012965:Sodium Chloride / Q000494:pharmacology*; D012995:Solubility; D000073893:Sugars / Q000378:metabolism; D014158:Transcription, Genetic
D018076:DNA, Complementary; D000073893:Sugars; D012965:Sodium Chloride; D010368:Pectins; D002482:Cellulose; D011392:Proline; D005964:Glucosyltransferases; C478648:cellulose synthase