Ueda, Katsumi

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RIGHT:URL:CITATION:AUTHOR(S):ISSUE DATE:TITLE:Structure of Plant Cells with SpecialReference to Lower Plants (II) :Feulgen's Nucleal Staining in SomeAlgaeUeda, KatsumiUeda, Katsumi. Structure of Plant Cells with Special Reference to Lower Plants (II) :Feulgen's Nucleal Staining in Some Algae. Memoirs of the College of Science, University ofKyoto. Series B 1956, 23(1): 87-911956-12-30http://hdl.handle.net/2433/258473ME,}foms oF THECOLLEGE OF SOIE,XTCE, UNIVERSITY OFVol. XXIII. No. 1, Article 11, 1956KyoTo. S)IRIEs B,Structure of Plant Cells with    Reference to Lower PlaRts.Feulgen's Nucleal Staining in SomeSpecialII.Algae '                                 By                            K.atsumi UEDA                           Cylolegical I.aboratory                    Botanical Institute, University of Kyolo.                          (Received 20 Oct. I956)    While many investigators have reported on Feulgen's ntic}eal staining inalgae, some of the results obtained by them do net aiways agree one another,esp:-cially in case of eyanophyta and conjugatae (cf. }<iesel and Doinikowa, 1937,Yamaha and Suematsu, l938, Niliary, l939). '    It is the purpose of the present investigation to employ the Feulgen stainingin various divisions ef a]gae wit-h a special attention to eyanophyta and conjugatae•Material and Methods    Nuclei of algae which belong to seven divisions were used for the material.IVEaterials were fixecl with ethanol, corrosive sublimate-ethanol mixture, or corrosivesublimate-acetic acid mixture, followed by wasking with water. After washing,they were hydrolyzed with AT-HCI in toto, or, if nece:-sary, in microtome sections,for 3, 5, IO, l5, and 20 minutes. After the hydrolyzed maaterials were stainedwith Sehiff's ureagent** frem 2 te 4, }.ours at room temperature (18-250C), theywere washed we}l with SO,-water, then were observed with a mieroscepe. Inmany species, however, the materials were sealed with canada balsam after de-iaydration and clearing.    Besides the nucleal staining, pyronin-methylgreen doub]e staining was carriedout to see the relation between t}ie nucleal staining and the double staining.ObservationsResu!ts of the ob$ervation are eeXNThi.s investigation was performedSchiff's Teagent was prepared bywithredqcinffpresented in the C.rant for  . "Meyck "the following table.Fundamental Seientific Research.basic fuchsin wit'n natriurp bisulfile,g88 Katsum{ [rEDASpeciesEEFixalives  Time of     .i OPtlMttml hydrolyslsEFeu]genstaining' Preparationsiascpa"ov=tc"pa pav "-" o m o v = se _ptOcrnywouaovrrtcrfpvscHnevcrg"paNostoc communeN. carneumN. piscina•LeOscitlatoria AnnaeO. tenuisEiO. IimosaO. princepsE,S-A,S-E   tl   rt   lt   x!tfltlny__sO. putridaCylindrosperntum  MichaiLovskoermseBrac}tytrichia balami.Gelidium elegansNemastoma       jTVIalcantieraeOrateloupta aptnisrl/1tlS-AllIS__555A-_nyA-- F+m totoCloiosiphonia       calm•ic}LaelCoraltin,a sp.in toto &mlcrotome sectionsWtirlii{510Synedra ulna(lyrosigna sp.CorMb'ella tumidiaC. aptnis        TSurireLZa eZegarbsS. tanceolataDitstrupt Brightwell'iPlanktoniella solBiddulphia stnensisCoscinodtsctts gigasIIIiIIII;I  S-A   ES--A, E  ES-A,S--E  S-A  E   tl   rl   ttDinophisis candinaCeratium hirunalinellaC. tTichocerosC. SitTcttC. fususIl:lfIE-A, E S-AS-E, F,  tl  r! 55 5555555le].ol5].515]5.m tototetraspoTemother celfs  .mlcrotomesect' ons ee-F '+I -mhi vegttativel cellsI in, totoISargassun} Horneri            egg            epid.S-A-l--1-++"}'-l--l'--- F+l5+""++- FÅ}+tn toto.tn toto .mlcrotomeSeCt!OnS Pyroninmethlgreen  stainingCells are stained diffusely, butnot differentially.Chromonemataand nuc}eoli arestained green andred respective{y, but the eolor--ilntenslty ls veryfaint.Chromonemataand nucieoli areweH difi'erentiallystained green andredi respeclively.Cltromonemataand nucleoli are differentiallystained green andred respect{ve}y.Ntrptoov=YptpoiiiltIiIItFl        StructureS. piLuliferum             egg             epid.S. Thunbergii             egg             epid.S. tortile             egg             epid.S. hemiphyttunt             egg             epid.Cystophyilum     StsyMbrioides             egg            epid.ffi•jikia fustfi rme             egg            epid.of Plant Cells with Speeia} ReferencePadlna arborescensColpornenia sinosalttttt  .lfltftlillilIiltiiilIIlIIIIII555555lifIEIfIÅ}+Å}+th+th+de+Å}+to LowerIlI'lllIEFP]ants. II.   iiitttl- 1 IO5.tn toto89mstOQ_o=ovrrtcntpoftcrp-ionev"w_paEu•gtena gracitisE. spirogyraFacu•s sp.CLosterium sp.Spirogyra setiformisS. sp.Zygnema sp.Micrasterias sp.NitetEa sp.          ;E,S-A,S-Ei 5          IS-A S-AS--A, E55s-A,E I S_tA I        I  tl  tt        ileEgg nuclei arediffusely stainedbtit chromonemataand nuc}eoli in acell of conceptacleepidermis aredifferentia}lystained.+++I in toto &I. mtcrotomejl sectionsac++rwNnt'f-l  .  vn totoin toto &mlcrotomeSeCtiOnS Min toto &mlcrotome   'sectlonsChromonemataand a nueleol"sare welldifferential}ystained green andred respectively. Abbreviatlons: • S-A; corrosive sublimate-acetic acid mixture. S-E; eorrosive sub}imate•ethanol mixlure. E; ethanol. epid.; conceptaele epidermi$. be Microtome sectienq. were sealed by eanada balsam after dehydration and clearing.    Besu}ts of the Feulgen staining in permanent preparaiions were not different from thQse    obtained by the observation immediately after washing with SOrwater.' It is seen in thiS table that the nuclei of whole species in cyanophyta andconjugatae were negative with the nucleal staining.     Among Feu]gen positive algae, it was observed in general that related speciestook nearly equal time of optimum hydro!ysis and were stained with similar toneand intensity, and t}aat chromonemata and nucleo]i were stained differentiallygreen and red respective]y when they were stained by methy]green and pyronin.   90 Katsumi UEDA      The nuclei of the algae be]onging to pyrropkyta were colered most intensely  with the F,eulgen staining, while these of rl}odophyta and phaeophyta were stained  very faintly, so fdr' as the material used in t}ie present study concerns.      Generally speaking, marine ehxysephyta seem to be stalned mere weak}y  than the $pecies in freshwater.                         Discussion and Conclusion      It has been reported by Peljansky and Petruschewsky (1929), Yamaha and  Suematsu (i938), Hillary (l939), and Drawert* (194•9), that cells of OsciZZatoria  are Feulgen positive. Other genera among cyanophyta have also been reported  that they show positive nucleal staining by Poljansky and Petru$chewsky (1929. )  in Spirttlina, Tolypothrix, Gloeotrichia, and by Bringmann (l950) in Lyngbya.  On the other hand, it has been stated by Kiesel and Deinikowa (1937) in Oscil-  Latoria and Totypotlwtos, by Herbst {.1953, l954,) in Oscillatoria, Aphanotheca,  Psendoanabaena, and by Shinke (unpublishecl) and Oura (unpublis}aed) in 0scil-  Latoria and other blue green algae that the cel}s are Feulgen negative. In the  present investigat-ion, it wes ebserved that many species of cyanophyta showed  negative nucleal staining in sectioned preparcations as well as in toto preparations.  Bringmann (1950) has repox#ecl that cells of OsciZZatoria are Feulgen positive  when the cells are treated wSth janthanum acetate before hyc]ro]ysis. In the  present investigatien, however, the cells ef OsciZlatoria arad Nostoc did not show  positive Feulgen staining though they were treated with Iantkanum acetate.• Besides cyanophyta, conjugatae is also a problematic group. For instance,  Geitler (l935), Yarnaha and Suematsu (1938), and Hiliary (1939) have reported  that tiie nuclei of Spi'rogyra are Fea]gen positive, w}}ile Shinke and Shigenaga  (l933), Kiesel ancl Doinikowa (1937) l)ave reported that Spirogyra is not stained  by the Feu]gen method. Besides Spirogyrct, Yamaha and Suematsu (l938) in  Zygnema and Hillary (1939) in CZosterium laave observed that these .algae show  positive nuc}eal staining. Four genera of conjugatae usec{ in tlte .present work,  however, showed negative nuc}eal staining. It must be noted here that the  nuclei of Spiregyra setbfornzis wa$ Feulgen negative throughout wkole mitotic  cycle in permanent preparatiens <Ueda, ln press).. Westbrook (l930, 1935), l<iese} and Doinikowa (1937), and Yamaha and  Suematsu (1938> in a few species of rhodophytca, Kiesel and Doinikowa (l937)  and Hillary (1939} in a few species of chrysophyta, Papenguss (1934i) ar}d l<iesel  and Doinikowa (1937) in a few species of phaeopliyta have reported that these  a]gae show positive nacleal staining. These results seem to be not disaccord  with those of the present investigation.       In egg nuclei of Sargassztm, the coloration was so faint that the present  * cited from Bringmann (1950)            Structure of ?lant Cells with Speeiai Referenee to Lower plants. II. 91author was not able to determine whether the Feulgen staining was positive er     .negatlve.                                  Summary     Feulgen's nucleal staining was' carried out in forty eiglit species among sevendivisions of algae.     Whole species of cyanophyta and conjugatae were Feulgen negative, alldother species were positive so far as the material used in the present investigatienconcemas.     Related species seem to take an egual time ef optimum hydrolysis and tostain similar tone and intensity.     Nueleoli were Feulgen negative in all species.     The author wishes to express his sincere tltanks to Professor N. Shinke forhis advice and eriticism through.out this work.                              Literature$ cited l) BRxNG}fANN, G., Planta 38: 54•1 (i950). 2) DRAvvERT, H., Planta 37: lel (].94•9). 3) GEirmeR, I.., Ber. Detitsch. Bot. Ges. 53 : 270 (1935). 4) llERBsT, F., Ber. Deutsch. Bo:. Ges. 66: 283 (I953). . 5> , Ber. Deutsch. Bot. Ges. 67; X83 (1954d). 6) H!LbARy, B.B., Bot. Gaz. 101:276 (1939). 7) KiEsEL, A. and DothiiKosvA, I'I., Cytologia Fujii Jnb. vol. pars ].: 10ti (1937). 8) I'ApEN"Fvss, G. G., Bot. Gaz. 96: 4•21 (1934,). 9) PollJANsKy, C. anG PETRvscl{EwsKy, G., ATch. Protis'. 67: II (}929).IO) SHiNKE, N. and SHrGENAGA, M., Cytologia 4i: l89 (l933).Il) WEsTBRooK, M.A, Ann. Bet. 4•4`:101! (1930).12)- ,Beih. Bot. Zentra!bl. 53: 564i (l935).I3) YArmHA, G. and SuEi[ATsv, S., Sei. Rep. TQIcyo Bunrika Dai.rr.aku. 3: 269 <1938).

Structure of Plant Cells with Special Reference to Lower Plants (II) : Feulgen's Nucleal Staining in Some Algae

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Structure of Plant Cells with Special Reference to Lower Plants (II) : Feulgen's Nucleal Staining in Some Algae

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