| In this study,through monitoring the changes of gene expression and physiological indexes in rice after different densities of BPH(brown planthopper)and N.cincticeps feeding for different lengths of time,we found that both BPH and N.cincticeps feeding stress all could effect the molecular and physiological of rice.The effect of rice Os CCT22,Os CAB1 R and Os10g0503200,the content of Chl and the activity of LOX in rice on low-density,short-term BPH feeding stress response were more significant.Protein kinase Os07g0537600,Os10g0503200,LHCB5 and Os SAUR24,the content of total protein and the activity of lipoxygenase in rice on low-density,shortterm N.cincticeps was more responsive to feeding stress.The detailed results are as follows:1.Effect of feeding stimulation on gene expression in riceAfter beening fed by BPH and black-tailed leafhopper,the RNA sequencing and transcriptome analysis of rice samples in each group were performed that there were 26 616 genes detected,of which more than 6 000 differentially expressed genes were present in each experimental group compared with the control group.After GO analysis of differentially expressed genes,there were 225 genes screened in the categories where the stimulus response was related to the response to stimulus;developmental process;positive regulation of biological process;negative regulation of biological process and growth.The absolute value of the expression ratio between the low-density group and the control group is greater than 2.5.eight of these genes are closely related to pests.2.The effect of BPH feeding stress on the expression level of defense-related genes in riceAfter beening fed by BPH,the relative expression levels of the monitored defense-related genes in rice all changed significantly.The expression of Os CCT22,OSCAB1 R and Os10g0503200 in rice have increased significantly were fed by BPH for 6 hours.The changes of Os07g0677400 was similar to Os07g0537600,they did not change significantly were fed by BPH for 6 hours,but the expression was significantly up-regulated of them were fed by BPH for 12 hours in rice.After beening fed by BPH with low density(1 BPH per plan),the expression of Os CCT22 in rice was 2.6 times higher than the control group(P<0.01).After beening fed for 6 hours by BPH with lower density(3 BPHs per plan),the expression of Os CCT22 in rice was 1.4 times higher than the control group(P<0.01).After beening fed for 6 hours by BPH with higher density(6 BPHs per plan),the expression of Os CCT22 in rice was 4.4 times higher than the control group(P<0.01).After beening fed for 6 hours by BPH with high density(9 BPHs per plan),the expression of Os CCT22 in rice was 1.2 times higher than the control group(P>0.05).After beening fed for 6 hours by BPH with low density,the expression of OSCAB1 R in rice was97.9 times higher than the control group(P < 0.01).After beening fed by BPH with lower density for 6 hours,the expression of OSCAB1 R in rice was 41 times higher than the control group(P < 0.01).After beening fed for 6 hours by BPH with high density,the expression of OSCAB1 R in rice was 2.2 times higher than the control group(P>0.05).After beening fed for 6 hours by BPH with high density the expression of OSCAB1 R in rice was 7 times higher than the control group 6 hours(P< 0.01).After beening fed for 6 hours by BPH with low density,the expression of Os10g0503200 in rice was 61% lower than the control group(P < 0.01).After beening fed for 6 hours by BPH with lower density,the expression of Os10g0503200 in rice was 2.7 times higher than the control group(P < 0.01).After beening fed for 6 hours by BPH with high density,the expression of Os10g0503200 in rice was1.9 times higher than the control group(P < 0.01).After beening fed for 6 hours by BPH with high density,the expression of Os10g0503200 in rice decreased 15%than the control group(P < 0.01).BPH feeding stress could activate the defense response in rice,the Os CCT22,OSCAB1 R and Os10g0503200 of rice pest related genes were strongly responsive on low density and short time feeding stress by BPH,and their expression levels changed significantly.3.Effects of BPH feeding stress on physiological indexes of riceThe contents of chlorophyll(Chl),total protein(TP),the activities of lipoxygenase(LOX),polyphenol oxidase(PPO),total superoxide dismutase(T-SOD)and peroxidase(POD)in rice were changed after beening fed by BPH.After beening fed with 2 BPHs per plan for 6 hours,the content of TP,the activity of PPO and LOX were significantly increased,the content of Chl was significantly decreased in rice.After beening fed with 4 BPHs per plan for 6 hours,the content of TP and the activity of POD were significantly increased in rice.After beening fed with8 BPHs per plan for 6 hours,the content of Chl,PPO and the activity of T-SOD were significantly decreased,and the content of Chl and TP,the activity of LOX were significantly increased in rice.After beening fed with 2 insect/plant density of BPH for 24 hours,the activities of LOX,T-SOD and POD increased significantly,while the content of Chl and the activity of PPO decreased significantly in rice.After beening fed with 4 insect/plant density of BPH for 24 hours,the content of TP increased significantly,the content of Chl,the activity of PPO and T-SOD decreased significantly in rice.After beening fed with 8 insect/plant density of BPH for 24 hours,the content of Chl increased significantly,the PPO,T-SOD and POD activities decreased significantly in rice.After beening fed with 2 insect/plant density of BPH for 48 hours,the content of Chl,the activity of T-SOD and POD were significantly decreased,while the content of TP was significantly increased in rice.After beening fed with 4insect/plant density of BPH for 48 hours,the content of Chl,the activity of PPO,T-SOD and POD and the content of TP were significantly decreased in rice.After beening fed with 8 insect/plant density of BPH for 48 hours,the content of Chl,the activity of PPO and T-SOD were significantly decreased in rice.After beening fed with 2 insect/plant density of BPH for 96 hours,the content of Chl,the activity of LOX and POD increased significantly,while the activity of T-SOD decreased significantly in rice.After beening fed with 4 insect/plant density of BPH for 96 hours,the content of Chl and the activity of T-SOD decreased significantly in rice.After beening fed with 8 insect/plant density of BPH for 96 hours,the content of Chl and the activity of T-SOD decreased significantly,the TP content and the activity of POD increased significantly in rice.4.Effects of N.cincticeps feeding stress on the expression of Defense-related genes in riceAfter beening fed by N.cincticeps,the relative expression of the monitored defense-related genes changed significantly.Os07g0537600 is the most sensitive to low density and short time feeding of N.cincticeps,while Os10g0503200,LHCB5 and OSSAUR24 are more sensitive to feeding of N.cincticeps.The expression of Os07g0537600 in rice after beening fed 6 hours by N.cincticeps with low density(1 insect per plan)was 2.28 times higher than the control group(P > 0.05).After beening fed 6 hours by N.cincticeps with low density(3insects per plan),the expression of Os07g0537600 in rice was 4.77 times higher than the control group(P<0.01).After beening fed 6 hours by N.cincticeps with higher density(6 insects per plan),the expression of Os07g0537600 in rice decreased by36% compared with the control group(P>0.05).After beening fed 6 hours by N.cincticeps with high density(9 insects per plan),the expression of Os07g0537600 in rice decreased 98%(P>0.05).After beening fed for 6 hours by N.cincticeps with low density,the expression of OS10G0503200 in rice was 54% lower than the control group(P>0.05).After beening fed for 6 hours by N.cincticeps with lower density,the expression of OS10G0503200 in rice was 84% lower than the control group(P>0.05).After beening fed 6 hours by N.cincticeps with higher density,the expression of OS10G0503200 in rice was 51% lower than the control group(P>0.05).Compared with the control,the expression of OS10G0503200 in rice after beening fed 6 hours by N.cincticeps with high density was significantly increased by 3.6 times(P <0.01).After beening fed 6 hours by N.cincticeps with low density,the expression of LHCB5 in rice was 99% lower than the control group(P > 0.05).After beening fed 6 hours by N.cincticeps with lower density,the expression of LHCB5 in rice was99% lower than the control group(P > 0.05).After beening fed for 6 hours by N.cincticeps with higher density,the expression of LHCB5 was 12.09 times higher than the control group(P < 0.01).After beening fed for 6 hours by N.cincticeps with high density,the expression of LHCB5 was 10.98 times than the control group(P<0.01).After beening fed for 6 hours by N.cincticeps with high density,the expression of LHCB5 in rice was 10.98 times higher than the control group(P<0.01).After beening fed for 6 hours by N.cincticeps with low density,the expression of OSSAUR24 in rice was 99% lower than the control group(P<0.0l).After beening fed for 6 hours by N.cincticeps with lower density,the expression of OSSAUR24 in rice was 99% lower than the control group(P<0.0l).After beening fed for 6 hours by N.cincticeps with higher density,the expression of OSSAUR24 in rice was 99% lower than the control group(P<0.0l).After beening fed for 6 hours by N.cincticeps with high density,the expression of OSSAUR24 in rice was 1.2 times higher than that of the control group(P>0.05).Beening fed by N.cincticeps,could activate the defense response of rice,and Os07g0537600,OS10G0503200,LHCB5 and OSSAUR24 of rice genes were tested by us had a strong response to low density and short time feeding stress from N.cincticeps,and its expression changed significantly.5.Effects of N.cincticeps feeding stress on physiological indexes of riceAfter being fed by N.cincticeps,the contents of Chl and TP and the activities of LOX,PPO,T-SOD and POD in rice were changed.After beening fed by N.cincticeps for 6 hours,the content of Chl of rice in all density groups,the activity of LOX in 4 insect/plant density group and the activity of TSOD of 4 insect/plant density group and 8 insect/plant density group were significantly lower than the control group in rice,while the content of TP of all density groups,the activity of LOX in 2 insects per plan density group and the activity of POD of 8 insects per plan density group were significantly higher than the control group in rice.After beening fed with 4 insects per plan t density group of N.cincticeps for24 hours,the content of total chlorophyll and chlorophyll b in 2 insects per plan density group,the content of TP,the activity of POD in 2 insects per plan density group,the activity of PPO of all density groups were significantly higher than the control group in rice,the activity of Lox in all density groups,the activity of T-SOD of 2 insects per plan and 4 insects per plan density group and the activity of POD in 8 insects per plan density group were significantly lower than the control group.The content of chlorophyll and TP,the activity of PPO in all density groups were significantly lower than the control group were fed by N.cincticeps with 8 heads/plant density for 48 hours,while the activity of LOX in 4 insects per plan density group and 8 insect/plant density group,the activity of T-SOD in 2 insects per plan and 8 insects per plan density group and the activity of POD in all density groups were significantly higher than the control group.Beening fed by N.cincticeps for 96 hours,the content of chlorophyll and TP,the activity of LOX,PPO and T-SOD in all density groups were significantly lower than the control group,while the activity of POD in 2 insects per plan density group and 4 insects per plan density group were significantly higher than the control group. |
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