Cloning And Expression Analysis Of Bell Family Gene In Actinidia Chinensis Planch.‘Hort16A'

Plant BELL family genes can regulate plant growth and development,such as controlling the transition of plants from vegetative growth to reproductive growth.Therefore,studying the potential role of BELL family genes in the development of kiwifruit flowers can provide a reference for the later regulation of kiwifruit flower development by molecular biology techniques.In this study,the phenological observation of flowering stage of kiwifruit,the division of flower development stage,total RNA extraction,identification of BELL family genes,screening and cloning,and expression analysis of target genes in different flower development stages,young leaves and young fruits of golden kiwifruit were studied.Obtained the following conclusions:1.By improving the reference method,a method for extracting total RNA from different tissues of gold kiwifruit was obtained.2.The annual flowering phenology,inflorescence and flower number,inflorescence location and inflorescence development of female and male plants of golden kiwifruit were very different.In addition,the flower development of gold kiwifruit was divided into 11 periods,named: 1: dormant bud period;2: bud expansion period;3: leaf emergence period;4: leaf extension period;5: current flower bud period;Flower bud stage;II: flower bud stage;III: 50% visible petal stage;IV: 90% visible petal stage;V: petal expansion period;VI: flower bloom period.3.9 BELL family members were identified from the kiwifruit genome database,including 22 sequences;some members have a unique domain of the BELL family,and some members have protein interaction and co-expression with genes involved in flower development,and they were differential expressions between different tissues.In addition,the family gene codon was weakly biased,and Arabidopsis was more suitable as an exogenous expression receptor for this family of genes.4.Five genes were obtained by homologous cloning,named AcBLH6,AcBLH7,AcBLH8,AcBLH9 and AcBLH11,and uploaded into GenBank.The accession numbers were MH667489,MH588307,MH667490,MH667491 and MH667492.Sequence analysis revealed that the AcBLH6 coding region contains 1947 bp,encoding 648 amino acids;the AcBLH7 coding region contains 957 bp,encoding 318 amino acids;the AcBLH8 coding region contains 2019 bp,encoding 672 amino acids;the AcBLH9 coding region contains 1695 bp,encoding 564 amino acids and the AcBLH11 coding region contains 1290 bp,encoding 429 amino acids.Evolutionary analysis found that the genetic relationship between the families was relatively close,and the homology between genes was high.In addition,AcBLH6,AcBLH8 and AcBLH9 all contain a BELL family-specific domain,but AcBLH7 lacks a partial BELL domain,AcBLH11 lacks VSLTLGL and an unknown domain.5.Expression analysis showed that the expression level of AcBLH6 gene was highest in young leaves;AcBLH7 gene was highest in 11 periods of flower development;AcBLH8 gene and AcBLH9 gene was highest in the first 9 periods of flower development.AcBLH11 gene was highest in the first 9 periods of flower development and in young leaves.