A T T I
DELLA
SOCIETÀ TOSCANA
DI
SCIENZE NATURALI
RESIDENTE IN PISA
MEMORIE - SERIE B
VOLo LXXXVI - ANNO 1979
ARTI GRAFICHE PACINI MARIOT!l - PISA - 1979
INDICE
DE DOMINIcIS V., CASINI S. - Memoria illustrativa per la carta della vege- tazione della Val di Farma (Colline Metallifere)
Explanatory notes on the Farma Valley (Colline Metallifere) vegeta-
tion map Pago
MICELI P., GARBARI F. - Cromosomi ed anatomia fogliare di quattro Allium diploidi di Grecia
Chromosomes and leaf anatomy of four diploid Allium of Grece . » 3 7 FERRI S., CAPRESI P. - Ricerche sui flavonoidi di Matricaria chamomilla L.
(Compositae)
Chemical investigation on Matricaria chamomilla flavonoids (Com- positae)
FERRI S., CARLOZZI C. - Influenza dell'idrolisi acida sulla morfologia, sulla cristallinità e sulla struttura dei granuli di amido
The effect of acid hydrolysis on the. morphology, the crystallinity and the structure of Potato starch grains .
CORSI G., PAGNI A. M. - Studi sulla flora e vegetazione del Monte Pisano (Toscana Nord-Occidentale). V. Le piante spontanee nella alimenta- zione popolane
Investigations on the flora and vegetation of Monte Pisano (North- Western Tuscany). V. The native plants in the human alimentation . VANNI S. - Note di erpetologia della Toscana: Salamandrina terdigitata,
Rana graeca, Coluber viridiflavus, Natrix natrix
Notes of erpetologia of the Tuscany: Salamandrina tergiditata, Rana graeca, Coluber viridiflavus, Natrix natrix
FAGOTTO F. - The Speke's Gazelle and its habitat in Somalia La Gazzella di Speke e i! suo ambiente in Somalia .
ONNIS A., STEFANI A., BISAIA L. - Ampelodesmos tenax Link (Gramineae):
leffietti della temperatura sulla germinazione in relazione alle condi- zioni dell'habitat
Ampelodesmos tenax (Gramineae): effects of temperature on germina~
» 53
» 63
» 79
» 103
» 125
tion in relation to habitat conditions . 133
MALLEGNI F., FORNACIARI G. - Su di un calvario turricefalo della tomba VII della Necropoli Eneolitica del Gaudo (Paestum)
A turricephalic calvarium of Burial VII in the Gaudo (Paestum) Eneo-
lithic Necropolis 149
BRANCONI S., DE DOMINIcIS V., BOSCAGLI A., BOLDI L. - La vegetazione dei terreni argillosi pliooenici della Toscana meridionale. I. Vegetazione pionilera ad « Artemisia cretacea »
Vegetation in the clayey Pliocenic soi! of Southern Tuscany. I. Pioneer
vegetation characterized by the presence of «Artemisia cretacea» » 163
VI
MALLEGNI F., FORNACIARI G., TARABELLA N. - Studio antropologico dei resti scheletrici della Necropoli dei Monterozzi (Tarquinia)
Anthropological study of skeletal remains of Necropolis of Monterozzi
(Tarquinia) 185
NAvARI-Izzo F., LOTTI G., GIULIANI P. M. - Ricerche sulle interazioni tra zin- co e acido gibber,ellico in Pisum sativum L.
Researches on the interations between zinc and gibberellic acid in
Pisum sativum L. . 223
RAIMONDO F. M. - Reperti per la flora briologica delle Alpi Apuane. Le rac- colte al Monte Procinto
Records for the bryological flora of the Apuan AI'ps. The collections
at Mount Procinto 237
CASSOLA F. - Un interessante reperto al Lago di Montepulciano (Siena): il Carabus clathratus antonellii Luigioni (Coleoptera Carabidae)
A noteworthy capture at the Lago di Montepulciano (Siena): Carabus clathratus antonellii Luigioni (Coleoptera Carabidae)
BALDERI F., TOMASELLI M. - Il paesaggio veg1etale della conca del Lago Tor- bido e del Lago Turchino al Monte Rondinaio (Appennino lucchese- modenese). III contributo. Nuovo reperto di Woodsia alpina (Bolton) S. F. Gray
Floristic and vegetational aspects of the glacial valley of Torbido and Turchino Lakes near Mount Rondinaio (Northern Apennines). III Con-
» 249
tribution. New record of Woodsia alpina (Bolton) S.F. Gray 253 ABBÀ G. - Flora esotica del Piemonte. Specie coltivate e spontaneizzate e
specie avventizie
Exotic flora of Piedmont. Naturalized and adventive species
SCRUGLI A., GRASSO M. P. - Contributo alla conoscenza delle Orchidaceae della Sardegna centrale
Contribution to the knowledge of Orchidaceae of CentraI Sardinia VERGNANO GAMBI O., PANCARO L., GABBRIELLI R. - Invlestigations on a nickel
accumulating plant: Alyssum bertolonii Desv. II. Phosphorus, potas- sium, iron and traCie element content and distribution during growth Ricerche su una pianta accumulatrice di nichel: Alyssum bertolonii Desv. II. Contenuto in fosforo, potassio, ferro e oligoelementi e loro distribuzione durante il ciclo vegetativo
CORSI G., MORELLI I., PAGNI A. M., CATALANO S. - Osservazioni morfologiche, isto-anatomiche, cario logiche e fitochimiche su Melissa officinalis s.l.
(Lamiaceae)
Morphological, histo-anatomical, caryological and phytochemical ob- servations about Melissa officinalis s.I. (Lamiaceae) .
MARCHIONNI V., ROLANDO A. - Influence of bonellin on the time of slex invler- sion and on fertility in Ophryotrocha pue,rilis
Influenza della bonellina sul momento della inversione del sesso e sulla
» 263
303
» 317
» 331
fertilità in Ophryotrocha puerilis 355
BRACALONI C., PISTOLESI G. - Indagini sulle zone Ulmide della Toscana. II. Il padule di Bientina
Investigations on the wetlands of Tuscany. II. Il «padule di Bientina» 363 TOMEI P. E., PISTOLESI G. - Indagini sulle zone umide della Toscana. III.
Aspetti floristici e vegetazionali del padule di Bientina. Nota preli- mina;J:1e
VII
Investigations on the wetlands of Tuscany. III. Floristic and vegeta-
tional aspects of «padule di Bientina ». A preliminary note 377 TOMEI P. E., ROMÈ A. - Indagini sulle zone umide della Toscana. IV. Con-
siderazioni sulle specie ornitiche fino ad oggi notle per il bacino del Bientina (Lucca-Pisa)
Investigations on the wetlands of Tuscany. IV. The birds of the ({ Pa-
dule di Bientina» (Lucca and Pisa districts) . 411 BARTELLETTI A., TOMEI P. E. - Indagini sulle zone umide della Toscana. V. Il
popolamento ornitico del Lago di Porta (Lucca, Massa-Carrara) Investigations on the wetlands of Tuscany. V. The birds of the ({ lago di Porta» (between Lucca and Massa-Carrara districts)
PAOLI G., PELOSINI I. -I gruppi sanguigni del sistema ABO negli scheletri di età romana di Collelongo (L'Aquila, Abruzzo)
ABO blood-group determination on Roman Age skeletons from Colle- longo necropolis (Abruzzo, ltaly)
PAGNI A. M., CORSI G. - Cariologia di alcune specie d'interesse officinale della flora italiana
Karyology of some species of ltalian officina l flora .
FICINI G., LUCCHESI G. - Sulla presenza dell'Aquila r:eale Aquila chry- saetus (L.) - in Toscana
On the presence of the eagle - Aquila chrysaetus (L.) - in Tuscany
» 433
» 459
» 465
» 475
Atti Soc. Tosc. Sci. Nat., Mem., Serie B, 86 (1979) pagg. 355-362, ff. 3, tabb. 3.
V. MARCHIONNI (*) and A. ROLANDO (*)
INFLUENCE OF BONELLIN ON THE TIME OF SEX INVERSION AND ON FERTILITY IN OPHRYOTROCHA PUERILIS
Riassunto - Influenza della bonellina sul momento della inversione del sesso e sulla fertilità in Ophryotrocha puerilis. Individui giovani di Ophryotrocha puerilis in fase maschile sono stati trattati con bonellina, il pigmento delle femmine mature di Bonellia viridis responsabile, secondo alcuni Autori, del viraggio della larva di Bonellia verso la mascolinità.
La bonellina ha rivlelato in Ophryotrocha effetto femminilizzante determinando un significativo anticipo della inversione alla fase femminile ed un incremento del tasso riproduttivo.
Summary - Young individuals of Ophryotrocha puerilis in male phase were treated with bonellin, the pigment of Bonellia viridis ripe flemales. The treatment induced a significant anticipation of the female phase and an increase of the reproductive rate.
Key words - Ophryotrocha puerilis - bonellin.
In Ophryotrocha puerilis, a proterandric hermaphrodite Po- lychaete worm, mutuaI influences between partners in female phase (<< Pair culture effect ») have been extensively investigated (HARTMANN and HUTH, 1936; BACCI, 1952; MULLER, 1962; PFANNEN- STIEL, 1975). Such effects lead to assume, in the mechanism of sex determination of this species, a regulating activity performed by the production of one or several pheromones.
In the course of experiments carried out in order to identify such substances, some assays have also been made by treating O.
puerilis with bonellin, a pigment which, according to some Authors (PELTER et al., 1976, 1978), should be «responsible for masculini- sation in the marine echiuroid Bonellia viridis ». This assumption, however, has never been demonstrated, because BALTZER and colI.
(*) Istituto di Zoologia dell'Università di Torino, Via Accademia Albertina, 17 - 10123 Torino.
356 MARooloNNr V. - RoLANDO A.
(1914-1937) and LEUTERT (1975) carried out their own experiments with total extracts, of which bonellin is only one component. Tne present attempt was suggested by the generally accepted idea that Echiurids are philogenetically related to Polychaetes and hence that there might be a similarity between sexually active substances in Bonellia and in Ophryotrocha. Moreover the multifactorial type of sex determination demonstrated in O. puerilis (BACCI, 1955) provides an interpretation for the existence of multiple sex phe- notypes also in Bonellia (BACCI, 1965; LEUTERT, 1975).
The experiments on Ophryotrocha have been carried out on the offspring of a single couple of animals collected at Mergellina (Bay of Naples, november 1977) and kept in standard conditions (at 18'°C, with naturallight and with spinach as food). The bonellin employed for treatment has been dissolved in seawater at the con- centration of 0.1 y Iml., since higher concentrations proved to be toxic. The medium has been partially changed twice a week in the course of the experimen ts.
Anticipation of the female phase. The animals were kept in stock cultures until they reached a lenght of 8-12 chaetigerous segments and were then isolated and subjected to treatment with bonellin, simultaneously control individuals of the same length were isolated. The evaluation of the moment of change from male to female phase and therefore of the degree of maleness or fema- leness of each individuaI was made - following Bacci's method
(1955) - by determining the number of chaetigerous segments shown at the time of the first appearance of free oocytes in the coelomic cavities.
The resul ts of the experimen ts show tha t bonellin induces an anticipation of the female phase of 0.62 segments in average, a difference which is highly significant (F = 8.11 + + +) (Table 1 and
TABLE 1 - The anticipation of the female phase by treatment with bonellin of isolated individuals.
rnean number
specirnens n of segrnents average an-
F a t sex change ticipation
treated with
bone11in 113 15.34 ± 0.16 0.62 8.11 +++
contro1s 113 15.96 ± 0.15
INFLUENCE OF BONELLIN ON THE TIME OF SEX INVERSION ETC. 357 Fig. 1). The daily individuaI growth was aiso considered and, as the animais were isoiated at five different Iengths ranging fra m 8 to 12 chaetigerous segments, the data were assembied in the five corresponding classes. The mean values show (Tabie 2) that the reduction of the growth rate with age is not different both in the treated and in controi specimens. This result Ieads to exclude that,
34 32 30 28 26 024 22
Cf)
Cl> 20
u c:: 18
Cl>
:::J 16
O"
Cl> 14
-
L.. 1210 8 6 4 2 O
Il treated N = 113
X = 15,34
D
controls: N = 113 X = 15,9610 11 12 13 14 15 16 17 18 19 20 21 number of chaetigerous segments
Fig. 1 -The specimens of o. puerili~ tnea,tyçl wit.q bonell~n anticipate the change te;>
the f(;!>male phas,e~ - 00 '
358 MARCHIONNI V. - ROLANDO A.
TABLE 2 - Average daily growths beginning from different size c1ass.
rnean dai1y growth from dif- da.i1y specimens n ferent size c1ass measuring average
in nurnber of segments growth
8 9 lO Il 12
treated with
bonel1in 113 0.94 0.77 0.70 0.68 0.66 0.75 contro1s 113 0.93 0.80 0.76 0.67 0.68 0.77
tota1 226
in the present instance, the bonellin may be regarded as a factor of growth.
Increase of the reproductive rate. 1) 24 animals, 12 specimens treated with bonellin and 12 controls, which inverted at the same Iength and in the same day, were kept isolated in order to controi the ripening of oocytes and the production of egg masses. The treatment with bonellin, carried on for about two months, has determined a remarkabie acceleration of the maturation of oocytes and the production of a greater number of egg masses as compared with controis (Tabie 3, Fig. 2 and Fig. 3). OnIy one among the specimens treated with bonellin did not Iay egg masses and showed resorption of the oocytes after a first phase of oocytes deveIopment
TABLE 3 - The increase of the reproductive rate by treatment with bonellin in female phase individuals kept isolated.
specimens number of number of egg n egg rnasses rnasses/individua1 treated with
bone11in 12 22 1.83
contro1s 12 12 1.00
tota1 24 34
INFLUENCE OF BONELLIN ON THE TIME OF SEX INVERSION ETC. 359 but as much as one third of the controls was unable to lay eggs.
2) 12 animals, 6 specimens treated with bonellin and 6 controls, which changed sex at the same number of segments and in the same day, were each mated with an individuaI in male phase (measuring lO chaetigerous segments) and kept under observation for a period of 40 days; the egg masses and the eggs number for egg mass of each pair were recorded. As shown in Table 4, the 6 pairs treated
TABLE 4 -The increase of the reproductive rate by treatment with bonellin in pairs.
pairs treated with bone11,in contro1s tota1
!Il Ol Ol Q)
2500
2000
'O 1500
L
.n Q)
È E 1000 ... Cl
... o
500
o
n
6 6 12
number of egg masses
2S 19 44
treated N = 7298
123456
numbet
eggsjpairjday of eggs
7298 32.87 4976 22.41 12274
controls N = 4976
6 5 4 3 2 1 number of coLiples
Fig. 2 - Reproductive rate: totai number of eggs spawned by each couple treated with oonellin and by eaçh couple of çontrol.
360 MARCHIONNI V. - ROLANDO A.
with bonellin laid a greater number of egg masses, a greater total number of eggs and hence showed a greater reproductive rate as expressed by the ratio number of eggs/day (Fig. 2 and Fig. 3) .
800
700
600
Cf) 500
Q)
u c
Q) 400
:::J
cr
Q)
L.. 300
...
200
100
. _ _ _ . treated ... controls
•...•..
..
3 5 7 9 11 13 15 17 19 21 23 25 egg masses
Fig. 3 - Number of eggs/egg masses in couples treated with bonellin and in couples of controI.
Parallel experiments were carri ed on with neobonellin (CA-
RIELLO et al., 1978), another component of the proboscis extracts of Bonellia, but no significant results were obtained.
IKFLUENCE OF BONELLIN ON THE TIME OF SEX INVERSION ETC. 361
The results of this work do not permit, at the present stage of the research, to put forward any assumption on the mechanism and on the specificity of the feminizing effect of bonellin on Ophryo- trocha. On the other hand, as mentioned above, experiments de- monstrating a specific masculinising action of bonellin on Bonellia larvae are completely lacking and we suggest that some features of bonellin point to it as un1ikely candidate to the role of sex deter- mining substance.
Especially: 1) bonellin is present in the integument of the whole body, not only of the proboscis, where the larvae adhere preferably, 2) bonellin seems to be the pigment responsible for the negative photobehavior of this Echiurid (LALLIER, 1955), 3) bonellin seems to have, as most integumental zoochromes, a chemical de- fence role owing to its high toxicity towards other marine animals
(MICHEL, 1931; LALLIER, 1955), 4) finally pheromones, in generaI, are not coloured.
The authors would like to thank G. Prota, L. Cariello, M. De Nicola Giudici and L. Zanetti (Istituto di Chimica Organica dell'Università and Stazione Zoologica.
Naples, Italy) for providing them with bonellin and neobonellin.
REFERENCES
BACCI G. (1952) - Diverso comportamento sessuale delle Ophryotrocha puerilis di Na- poli e Plymouth. BolI. Soc. Biol. Sper., 28, 4, 1-4.
BACCI G. (1955) - La variabilità dei genotipi sessuali negli animali ermafroditi. Pubbl.
Staz. Zool. Napoli, 26, 110-137.
BACCI G. (1965) - Sex determination and sex balance of Ophryotrocha puerilis. Nature, 207, 1208-1209.
BALTZER F. (1937) - Entwicklungsmechanische Untersuchungen an Bonellia viridis.
III. Ueber die Entwicklung und Bestimmung des Gesch1echts und die Anwend- barkeit des Goldschmidtschen Zeitgesetzes der Intersexualit1:it bei Bonellia viri- dis. Pubbl. Staz. Zool. Napoli, 16, 89-159.
CARIELLO L., DE NICOLA GruDICI M., ZANETTI L., PROTA G. (1978) - Neobonellin a new bio- logically active pigment from Bonellia viridis. Experientia, 34, 1427-1429.
HARTMANN M., HUTH W. (1936) - Untersuchungen ii. Geschlechtsbestimmung ii. Gesch- lechtsumwandlung von Ophryotrocha puerilis. Zool. Jahrb., 56, 389-439.
LALLIER R. (1955) - Recherches sur la toxicité des extraits du V·er marin Bonellia viri- dis. c.R. Acad. Sci., Paris, 240, 1489-1491.
LEUTERT R. (1975) - Sex determination in Bonellia. In, Intersexuality in the AnimaI Kingdom. Reinboth (ed.). Springer Verlag, Berlin.
MICHEL F. (1931) - Ueber den chemischen Schutz der Bonellia viridis gegen Fra,ss.
Pubbl, Staz. Zool. Napoli, 11, 22-29.
362 MARCHIONNI V. - ROLANDO A.
MULLER H. (1962) - Deber die sexualitat des PoIychaeten Ophryotrocha .puerilis, ihre Determination und ihren Einfluss auf Driisentatigkeit KauapparatentwickIung.
Z. Morph. Okol. Tiere, 52, 1-32.
PELrER A., BALLANTINE J. A., FERRITI V., JACCARINI V., PSAILA A. F., SCHEMBRI P. J. (1976) - Bonellin, a most unusuai Chiorin. J. Chem. Soc. Chem. Commun., 999.
PELrER A., BALLANTINE J. A., MURRAY-Rusr P., FERRITO V., PSAILA A. F. (1978) - The structures of anhydrobonellin and bonellin, the physiologically active pigment from the marine echiuroid Bonellia viridis. Tetrahedron Letters, 21, 1881-1884.
PFANNENSTIEL H. D. (1975) - MutuaI influenoe on the Sexuai Differentiation in the Protandric PoIychaete Ophryotrocha puerilis. In, Intersexuality in the AnimaI Kingdom. Reinboth ed. Springer VerIag, Berlin.
(ms. preso il 18 dicembre 1979; ult. bozze il 18 giugno 1980).
