DNA CONTENT IN SPECIES BELONGING TO THE FAMILIES ACRIDIDAE, OMEXECHIDAE, ROMALEIDAE (SUPER-FAMILY ACRIDOIDEA) AND PROSCOPIIDAE (SUPER-FAM[LY PROSCOPOIDEA) # AMILTON FERREIRA Depí:%:g£adeF.£#SUEgS,"]S3í.d5aod§,ERS;oadsu,%'rop,ag#.S#r;:Jí,úti° H"ODUCTION There .are ín the ]íterature few Papers Concerneá Wíth the DNA contept ín the orthopteroid insects. One of the work on this subieét come from John and Hewitt (1966). According to them, the karyotypic stability assumed for the species of the family Acrididae in studies made on chromosome number and structure is not true when the amount of DNA of spermatid nuclei are t-aken on accoun£. In this opportunity is our intention to study the DNA content of the followings spec.\es.. Rbammatocerus conspersus, Metal_eptea brevicornis, Xyleus sP, Aleuas gracilis, Omexecba servillei and Cepbalocoema borelli. Rbammatocerus conspersus, Metaleptea brevicor&is and Xyleus sP., tiLkev!.i`s`.e dm majority of species belonging to the family Acrididae have a karyotype with 2n (ó) = 23 acrocentric chromosome that is considered basic for the Cryptossacci. In A/cz/¢s g"!c?./z.s Lh:ío°ncct::£n;fac:fbte¥:e::n*Cofauusí:::mhe¥::dtíf:e:thúSírsbbe*jceekna?h:¥epxe¡ch°r:ei::ot±: hmüdcahna:;::s:h=ek:hteó;í;rktxozS,ga;eexr:seec£:naís]:rgte¿¥e+te¥óe#¥L:Óa,u;o;:.mAesaancdo¥s:á:::::, the diploid chromosome number was reduced from a 2n (o) =23 to 2n (Ó) =20 (Ferreira, 19?.4, 1975). 0. seiiJ¢.//e¡. has a karyotype with 2n (6) = 23 (Piza,1952; Saez,1956;Mesa, 1956 e 1963; Ferreira,1974). Save the largest pair of autosomes that is metacentric, all the others, even the X are acrocentric. Cepb¢/ococ77M bore//g. has 2n (@ = 19 acrocentric chromosomes (Ferreira. 1974). With the data here obtained the DNA content of species belonging to different families, as well as the DNA content of species belonging to the family Acrididae may be compared. *cTohj:eyho:kN£Ponns3ftd:3XeFnuv:?vaig£Pendt:ái=nptaírf?coáepiscqc:i;?óá:oE(sct#q,?esáopaulo(FAPESP)md REV. SOC. ENT. ARGENTINA, Tomo 36 (1-4): 35-40. 36 Revista de la Sociedad Entomológica Argentina 36,1977 MATERIAL AND METHODS The species here studied as well as their collecting localities are listed below. i. Super family Acridoidea. Family Acrididac Sub family Copiocerinae A/cc#s gr#cz./í.s Stál -Corguinhos, Mato Grosso, Brasil. Sub family Gomphoccrinae Jtzm7%772¢foc'erz# c'o77spe7isz# (Brunner) -Corguinhos, Mato Grosso, Brasil. Sub family Acridinae Me£c}/ep£c¢ Z777.¢c.co77%.s (Blanchard) -Corguinhos, Mato Grosso, Brasil. Family Romaleidae Sub family Romaleinae Xy/ez£s sP (Ilha Solteira, Sáo Paulo, Brasil). Family Omexechi'dae 0777c#ccb¢ sg7ÜÍ.//cí. (Blanchard) - Rio Claro, Sáo Paulo, Bra.sil. \ 2. Super family Proscopoidea \ Family Proscopiidae ``1+ Ccpb¢/ococ777¢ bo7'c% (Giglio-Tos) -Corguinhos, Mato Grosso, Brasil. SPÉ7w7?¢£¢.ds 7?z/c/cg. were chosen for the DNA measurement because they are easily identified md have the chromosome material uniformily distributed in it. The testes removed from de males were cleaned of the investing fat body ahd fixed for 12 hours in Carnoy 3: 1. The fixed material was hydrolysed in NHcl at 60.0 C during s minutes and transfered to Feulgen reagent for one and a half hour. The Feulgenized material was bathed in tap water and finally lightly squashed® The testes of all the species received the same treatment, i, e, fixation, hydrolyse and staining were done at the same time aiid with the same stock solution. The DNA content per nucleus in arbitrary units wa; determined according to Frazer and Davidson (1953). All the extintion~-values were taken in a Zeiss citophotometer at the wavelength of 560 nm fo`i each of 10 nuclei from 3 individuals in each specie. The extintion value for each nucleus was the mean of three values obtained at three different plugs. The values of the DNA content obtained for each species were arranged in histograms (Fig.1). A~ FERREIRA,.DNA contents in Acrididae 37 i-Omexecha servil!ei 2-MetQleQL± brevicornis 3-E¥Lbs ip 4-Aleuas _ _-___ - racilis 5-Rhammatocerus conspersus DNA-Value (Arbitrciry units} 6-Cephaiocoema boreHi Fig.1 -The DNA content (in arbitrary units) of lc spermatid nuclei in 6 species of orthoptera (3 species of ACTididae, 1 specie of Romaleidae), 1 specie of Omexechidae and one of Proscopiidae). E ach histogram is based on 10 nuclei from each 3 individuals. The total number of nuclei measured per species is 30. 38 Revista de la sociedad Entomológica Argentina 36,1977 RESULTS d9 The analysis of variance of the DNA content (Table 1) obtained photometrically in spermatids nuclei shows that no significant differences exist between individuals of the same specie but significant differences were observed between species belonging either to the same, or to different families. TABLE i. Analysis ofvariance of the DNA content in 5 Pecies of the supe-r Family Ácridoidea ®e|on- Éng to three differents famiHcs) and onc of the Supcr-Family-ProscopoideaL Source ofVariation df SS MS F -15o007. 6760 Specie 5 3001.5352 1762.1882 * Individuals inthespecie 12 20.44 1.7033 0.3185 n.s. `-86.664.830 Residual 62 5.3487 Total 0.05 ` 15.894.5990 * = significant n.s. = no significant at 0.05 The-Tukey test (Table 2) shows that there are s:Énificant differencés in the DNA content in most of the species. Howewer no significant differences were observeit between Omexecba servillei, Metaleptea brevicornis a;nd Xyleus sP a;nd a.1so between Aleuas gracüis a.nd Rbammatoc,erus conspersus. TABLE 2. Tukey test aplied to cytophotomctric dari`i b-ars conect species not differing significantly at the 50/o level. Specie Family XDNA Omexecba servillei Omexechidae 41.39 Metaleptea brevic ornis Acrididae 42.03 Xyleus sp Romaleidae 3 9. 26 Aleuas gracilis Acrididae 49.00 Rbammatocerus conspersus Acrididae 47.03 Cepbalocoema borelli Proscopiidae 2 0. 9 3 Only two of the species of Acrididae with similar karyotypes show no significant difference in DNA content (Table 2). AL. FEE+REIRAL, DNA contents in Acrididae 39 DISCUSSION The results of DNA content obtained cytophotometrica]ly in species of the family Acrididae with similar karyotypes show that chromosomal rearrangement not detetable at chromosomal level ocurred in these species during the philogenetic process, since significant differences in DNA content was observed between them. Similar results were obtained by John and Hewitt (ig6ó).iThe nature of these chromosomal rearrangement is unknown at the moment, but the most probable hypotesis is concerned with the occurrence of duplication or dificiency (Rees and Jones,1967). In spite of the great chromosomal rearrangement that has modified its karyotype (Ferrcira, 1974, 1975) A/cG£¢s gy¢w./í.s has a DNA content not significant different from Rbcz77c77?¢£occrz/s co7¢spcr.s#s that has a karyotype without any aparent change in its organization® The similarity of DNA content observed in 077?c#ecb¢ scrt;z.//cÍ. in Xy/cc# sP iand in Rbcz77277?¢£occ7"s co7?s,')crs#s seems to shows a close phylogenetic relationship between the families Acrididae and Omexechidae thought more species should be studied. This point of view is in according with the cytological data. Mesa (1963, 1964) has claimed that the basic karyotype of the family Omexechidae is secondarely derived from the 23 acrocentric chromosomes normally found in the species of the family Acrididae by a pericentric inversion in its longes+¿ autosomes. Cepb¢/ococ77?cz borc//z., the single species of the family Proscopiidae here studied, has approximately half of the averaged DNA content found in the remaining five species, which belongs to the families Ommexechidae (1 sp), Acrididae (3 sp) and Rommaleidae (1 sp). Thought fragmentaries, sinc€ more species shoul be taken on account in future studies our observations seems to show no close relationships between Proscopiidae and the above mentioned families. Also from the citological point of view the species óf the family ProscQpiidae depart from the classical 2n= 23 -24 of the true grasshoppers. In fact, according to several papers published (Piza,1943,1945; De Castro,1946; Dasgupta, 1960; Santiago, 1968; Mesa, 1973; Ferreira, 1974), de most common number in Proscopiidae is 2n = 19-20, with all the chromos()mes acrocent..'ic. In a few species (Mesa, 1973 and Ferreira, 1974) this karyotype has been modified throught several structural rearragements. The above mentioned observations agree with the recent point of view of Kevan (1966') and Blackith and Blackith (19661967 a and 1967 b), that based on morphologiéal considerations removed Proscopiidae from Acridoidea. The authors placed it on Eumastacoidea and recently Descamps (1973) studyng the phallic complex of species belonring to 9 genus from the 14 álready known creat.ed the Super-family Proscopoidea with a single family Proscopiidae that possess all the species described. SuhmRy !Fe#,fchá:;P:tiaí:?!T,t(Í,:e::ceo:jiioííe¥wr:¢rseRge:e::l'iqn?|ícm:t?;e'íoíctceobáses:cn;cZsíÍ;zísf::=T:Í:a:'eFPse;Íe:rcf;Ía:e:,:#n'á The results obtained have shown mainly that: a) species with the same karyotype may have #fi{ch¥:affí::ehcá:d:;p`9teheLENR£Í:[ne:3::;2dt:,et£g¥;[Lfymper%::3];dí]Fdea:e£gasoprboexícLoastee][yy#fteodf the DNA content found in the species of the families Omexec!`idae, Acrididae and Romaleidae. REFEI(ENCES BLACKITH, R. 8. and R. M. BLACKITH (1966)-The anatomy and physiology of the Morabine qrasshoppers.1. Digestive and reproductive svstems. Aus. .T. Zool.14: 31-40. 40 Revista de la sociedad Entomológica Argentina 36,1977 BLACKIT:ássRrio3bear:.d|F..#:te¥±?cgaTOTm(ylg£7dac)o-ml;haensgnastowTtyh¥ydrgpohí3iropl£gaeo,fÁt:dTdoareabiná Proscopiidae. Aust. J.