Study On Developmental Genes Related To Hindleg Specialization Of Atrijuglans Hetaohei Yang

Atrijuglans hetaohei Yang is the main fruit pest on Juglans regia L,an important woody oil tree in China.compared with general moths,the structure and behavior of hindleg are different.when crawling or standing on the surface of objects,the hind leg lift and swing to both sides of the body.at present,there is no research report on the morphological characteristics and behavior of hind leg of adult Atrijuglans hetaohei Yang.In this study,the ultrastructural changes of the joint parts of the chest and leg of 3 Juglans regia were observed by scanning electron microscopy,and the transcriptome of the chest and leg of Atrijuglans hetaohei Yang was sequenced by high-throughput sequencing platform,and the genes that may be related to the specialization of its hind leg were screened.The main research results are as follows.(1)By sequencing and analyzing the transcriptome of different genders’ fore leg tissues,a total of 86.11 Gb clean data was obtained,and 186,361 transcripts were obtained by splicing optimization,with an average length of 936.04 bp and an N50 value of 1496.By comparing and analyzing the transcriptome data,a total of 59 genes related to the specialized development of hind leg of Atrijuglans hetaohei Yang were screened out.Among them,Including a decapentaplegic(dpp),a Distal-less(dll),two homothorax(hth),a extradenticle(exd),two apterous(ap),a Antennapedia(Antp),a Sex combs reduced(Scr),a empty spiracles(ems),a engrailed(en),two vestigial(vg),a abdominal-A(abd-A),a abdominal-B(abd-B),a Ultrabithorax(Ubx)and 43 other homeobox genes(home obox genes,hox)(2)Seven candidate genes including Ubx,Dll,dpp,exd,hth,en,abd-A were analyzed by RT-qPCR,and the results were basically consistent with the change trend of transcriptome sequencing analysis.The expression of seven genes was different in different developmental stages,adult stage > larval stage > pupal stage.In pupal stage,the expression levels of seven genes in hindleg were significantly lower than those in foreleg and midleg,except that the expression levels of en and dpp in midleg were higher than those in foreleg.Combined with relevant literature reports,it is speculated that these six genes are closely related to the development of chest and leg of Atrijuglans hetaohei Yang,and Ubx gene promotes the specialization of adult hindleg,leading to the specialization of hindleg.(3)Scanning electron microscope was used to observe the ultrastructure of the front,middle and rear leg of Atrijuglans hetaohei Yang.Compared with the front and middle leg,the hind legs were enlarged and setae were developed,but the types and distribution of sensilla on the three pairs of chest leg were basically the same.By comparing the connection modes of walnut limb lifting moth 3 to each joint of chest and leg,it was found that the anterior leg trochanter was kidney-shaped,which was connected with the protruding basal ganglia from the inside,and the connection was embedded with two bony processes The middle leg trochanter is bow-shaped,and when connected with the basal ganglia,the two protrusions of the trochanter are stuck in the two joint seams formed by the four bony processes of the basal ganglia;The hindleg trochanter is sector,which is connected with the basal ganglia by one corner end.There is an arc-shaped protrusion on the basal ganglia,and the smaller trochanter just buckles on the basal ganglia protrusion.Combined with relevant literature reports,it is speculated that the fan-shaped shape of the hindleg trochanter of Atrijuglans hetaohei Yang and the arc-shaped protrusion structure on the basal ganglia provide support for its hindleg.In this study,Ubx,exd and other candidate genes related to the specialized development of hind leg of Atrijuglans hetaohei Yang were preliminarily screened by transcriptome sequencing and RT-qPCR technology,and the structural differences between the base of hind leg and the front and middle leg were made clear by scanning electron microscopy,which provided a research basis for further studying the biological characteristics of lifting and swinging of hind leg of Atrijuglans hetaohei Yang.