Analysis Of Tobacco JAZ Gene Family And Gene Function Of NtJAZ10

JAs are not only widely involved in regulating plant growth and development,such as seed germination,root growth,flower development,tuber formation,pigment synthesis,fruit maturation and senescence,but also play an important regulatory role in biotic and abiotic stress responses.The JAZ protein is a key inhibitor of the JAs pathway.In order to systematically analyze the basic characteristics of the common tobacco JAZ gene family and the function of the NtJAZ10,this study first identified 23 NtJAZ from the common tobacco genome,and then systematically analyzed the NtJAZ gene family’s phylogeny,gene structure,conservative motif,chromosome location,repetitiveness,tissue expression characteristics,and abiotic stress and hormone response characteristics.On this basis,we cloned the NtJAZ10,performed overexpression analysis and knockout analysis,respectively,to identify the role of NtJAZ10 in tobacco growth and development.The main findings are as follows:1.23 kinds of NtJAZ gene sequences were identified from the common tobacco genome database,all of which contained the conserved TIFY and Jas domains.The length of the NtJAZ coding sequence is between 303 and 206 bp,the length of the amino acid sequence is between 101 and 402,the molecular weight is low,and the size is between 11.65 and 42.62 kDa,the number of exons of NtJAZ ranges from 2 to 8,and the NtJAZ protein Rich in basic amino acids.Collinearity analysis showed that there were three pairs of gene fragment duplications in 23 NtJAZ gene sequences.Evolutionary analysis shows that tobacco and tomato JAZ are highly similar.2.Analysis of tissue expression characteristics found that the expression patterns of 15 NtJAZ genes in different tissues are different.NtJA2-4,NtJA2-14 and NtJAZ-15 have no obvious tissue differences in expression.NtJAZ-1 is found in cotyledons,seeds,High expression in roots and peeled stems.Compared with other tissues,NtJA2-2,NtJAZ-3,NtJAZ-5 and NtJAZ-10 are expressed at high levels in roots,and NtJA2-7,NtJA2-8,NtJAZ-12,NtJAZ-13 in cotyledons the expression is weak,while NtJAZ-11 is only expressed in cotyledons.The transcription levels of NtJAZ-3,NtJAZ-5,NtJAZ-9 and in the stem epidermis are higher than those in the debarked stem,indicating These four NtJAZs may be particularly related to epidermal development.The 15 NtJAZ genes showed different expression patterns under abiotic(high salt,drought,cold,high temperature)stress and different hormone treatments(SA,ABA,MeJA,GA).Under high salinity(NaCl)treatment,the expression of all NtJAZ genes are up-regulated;under drought(PEG)stress,NtJAZ-1,-2,-3,-4,-5,-6,-7 and-8 The expression of eight NtJAZ genes was up-regulated,and the expression levels of the remaining genes were suppressed by drought stress;under cold(4℃)stress,all NtJAZ genes were activated;compared with other stresses,all NtJAZ genes were activated at high temperature(42℃).The expression level of the NtJAZ gene increased.After SA treatment,the expression of most NtJAZs genes was significantly increased,while NtJAZ-1 was only slightly upregulated;after ABA treatment,except for the non-activated NtJAZ-1,-2 and-3,NtJAZ-4,-5 and-6 were upregulated.The expression levels of these three genes inhibited the expression of other NtJAZ genes;through MeJA treatment,the expression of ten NtJAZ genes was activated.After GA treatment,the transcription of NtJAZ-5 was inhibited,while the expression of other NtJAZs increased.3.Clone the NtJAZ10 gene of common tobacco,and analyze the gene overexpression and knockout respectively.Firstly,the overexpression vector pCAMBIA-NPT-GUS-NtJAZ10 and the gene editing vector NtJAZ10-knockout were constructed to infect tobacco K326 respectively to obtain NtJAZ10 gene overexpression plants(K326-T)and NtJAZ10 gene knockout plants(K326-M).Through identification and phenotypic testing of transgenic plants,it was found that K326-T plants had no stamens and the ovary changed from 2 carpels to 3 carpels;while the stamens and ovary of K326-M plants were basically normal,but the stigma hairs were missing;NtJAZ10 overexpression and knockout lines were aborted and could not be fertilized to obtain seeds.The above results prove that JAZ-10 has a key regulatory effect on the development of plant reproductive organs,and it has important application value in the construction or screening of seed abortion plant varieties.