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Maria Neysa Silva Stort and Elza Aparecida Dos Santos Pavanelli

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Maria Neysa Silva Stort and Elza Aparecida Dos Santos Pavanelli Rev. Biol. Trop. 34 (1): 59-62, 1986 Crossing systems in Ep idendrum nocturnum Jacq. (Orchidaceae) Maria Neysa Silva Stort and Elza Aparecida dos Santos Pavanelli. Departamento de Biología, Instituto de Biocieneias, UNESP 13.500. Rio Claro Brasil. (Reeeived: April l9. 1985) Abstraet: Unter greenhouse eondictions Epidendru m nocturnum (Orehidaeeae) produced many fr uits by seU-pollination and clei5togamy. These processe5 must be optional for the group, sinee intra- and interspeeifie pollinations abo lead to the formation of fr uih having seed5with embryos. The seedsproduced by different erossing systems were either without embryos or had 1 to 4. Extra embryos appear more frequently in fruits formed without artificial pollination. On the average, the seeds obtained by artificial eross-pollination germinated more than those removed from naturally formed fruits. Most orchid species reproduce by allogamy Stort and Martins (1980), in a study of the and their flowers showadaptations to cross-pol­ genus Cattleya, observed that self-pollination lination. According to Brieger (1966), the con­ leads to decreased production of seeds with em­ stitution of the pollen masses of orchids, as well bryos. Of the fifteen species studied, only e as the presence of the rostellum, a structure aurantiaca, which normally reproduces by self­ that separates the anther from the cavity of the pollination, showed higher mean percentages of stigma, cause orchids to be dependent on ani­ seeds with embryos when pollinated artificially. mals fo r pollination. According to Dodson In the present study we attempted to deter­ (1955), about 80% of all orchid species are pol­ mine the reproduction systems of E. noc­ linated by insects, 3% by hummingbirds, and tumum under greenhouse condictions; we sur­ 15% by diffe rent pollinating agents. Only 20/< veyed the production of seeds with and without are self-pollinating. embryos and determined the germinating power Self-pollination may be an optional or obli­ of the seeds produced. The data were compared gate reproductive phenomenon among orchids. with those obtained by artificial (manual) pol­ Thus, sorne species that normally reproduce by lination. cross-pollination can be self-pollinated in the absence of their natural pollinating agents MATERIAL AND METHODS (Hagerup, 1952). Other species are deistoga­ mous, i. e, they are self pollinated before the The plants of this study were field-collected flowers open. Reports of self-pollination among in Costa Rica and Panama, and kept under orchids have been published by Standley (1928), greenhouse conditions fo r several years. On the Hagerup (1952), Kirchner (1952) and Knudson basis of anaIysis of the morphological traits and (1956). distributioh of different E. noctumum popula­ 1 A few species of orchids reproduce by deis­ tions, Brie er & Bicalho (1978) subdivided this togamy, as observed by Darwin (1877), Rein­ species int groups which the caBed: E trídens chenbach (1977), Forbes (1885), Smith (1905 ; Poepp. et �nd\., E. oliganthum Schltr., E. ba­ 1928; 1929), Schlechter (1914), Swamy híense Re chb . and E. noctumum var. tagua­ (1949), Kirchner (1952), and Ilgg (1975). tíngense B ieger e Bicalho and E. erectu m. In Contrary to what occurs in most orchid spe­ our study we utilized plants of the groups E. des, which, in greenhouses do not produce tridens, E. noctumum var, taguatíngense and fruits, E. noctumum specimens produce many E. ere�tu�. Her� we will consider the �roups fruits practically throughout the year. as a Single specles (E. noctumum), as It was 59 60 REVISTA DE BIOLOGIA TROPICAL originally considered, and based on the results TABLE l of experimental hybridizations by Pavanelli and Stort (1985). Percentage average 01 seeds with em bryo After the flowers opened we kept records of obtained by natural and artificial pollination visitors to the flowers to determine the pres­ sence of pollinating agents. Also, we tried to Artificial pollinations verify the reproductive systems of the plants. Species Natural For seed observations we collected ripe fruits pollination Self- Intraspecific obtained by natural pollination (i .e. formed E. nocturnum 59.43 20.92 48.74 without human intervention), and fruits formed E. erectum 28.52 22.28 46.28 by artificial (intra- and interspecific) pollination E. tridens 44.55 • 71.53 and by self-pollina tion. Artificial pollination Mean 44.16 21.60 55.52 • was performed manually by transferring a pol­ No self- pollination linarum of one flower to the stigmatic surface The percentages of seeds with and without of the same (self-pollination) or another flower embryos and seeds with different numbers of (cross-pollination). Crosses were performed embryos varied according to the type of polli­ between plants from the same region and be­ nation performed and to the plants involved in tween plants from different regions, with a the process. In view of the large number of data total of 218 pollinations. A total of 31 self-pol­ obtained with the different pollination systems, linations were performed. Five seed samples we present the mean values. Table 1 shows the were removed from each fruit and counted by percentages of seeds with embryos found in the tecnique utilized in previous studies (Stort, fruits formed after natural pollination (includ­ 1970). ing self- and cleistogarny) and after artificial Seedswere tested fo r germination by sowing self-and intraspecific pollination. samples from each fruit in culture media accord­ The E. nocturnu m plants showed higher ing to Knudson (1946). Two months after percentages of seeds with embryos in naturally sowing, 10 sarnples were removed from each formed fruits. In E. erectum and E. tridens medium and 10 slides were prepared. Nine higher percentages were obtained for fruits microscopic fields were counted on each slide. formed after artificial intraspecific cross-polli­ The results were compared by the non-pararnet­ nation. ric Kruskal-Wallis test and by the Mann-Whitney Considering the group as a whole (pavanelli Uand Z tests, according to sample size. and Stort, 1985), a higher percentage value was obtained for fruits after artificial intraspecific RESULTS pollination (55,52%) than afier self-pollination (Z = 3.79, p<0,05). The mean percentage of Self-pollination and cleistogarny are the nat­ seeds with embryos found after natural polli­ ural processes that lead to fruit fo rmation in nation (44.l6%) was higher than that obtained the plants studied under greenhouse conditions. by artificial self-pollination (2 1 .60%). StatisticlJl No pollinating agent was ever observed on these comparison of these results yielded U = 6.00 flowers throughout five years. We noted that (significant , P<0,05). On the other hand, the the rostellum is more developed in the lateral percentage average of seeds with embryos ob­ than in the central portions. This type of fo r­ tained in intraspecific (5 5.52%) and in inter­ mation facilitates self-pollination. A few days pecific crosses (57.31%) involving the species after opening of the flower, thepollinia contact E. nocturnum and E. erectum are not statisti­ the stigma through the central portion of the ca1ly significant (Z=0.48). SeeTable 1. rostellum that degenerates as the polien tubes fo rm , while the walls of the colurnn grow to­ Numbers of embryos in the seeds. E. noctur­ wards the central portion, closing it, and the pet­ num seeds ow varying numbers of embryos, als and sepals wither and the ovary develops both as previousl indicated. By considering only the intemally and extemally. Intemally, megasporo­ seeds with e� bryos, the following results were genesis normally occurs, leading to ovule fo rma­ obtained f0 naturallY formed fruits: 68.92% tion. The seeds fo rmed either had no embryo or seeds with o e embryo, 26.36% with 2, 4.35% only 1 to 4 embryos. The embryos are of sexual with 3 and 1.37% seeds with 4 embryos. In origin or apomictic(Stort and Pavaneli, 1985). seeds forme after artificial intraspecific polli- SILVA & PA VANELL/: Crossing systems in Ep idendrum 61 nation we found: 91.07% seeds with one latter exhibit, on the average , higher numbers embryo, 8.07% with 2,0.85%with 3and O.0 1% of aditionad embryos, which besides being with 4 embryos. Thus, the frequency of 0.01% smal1er than single embryos and therefore less extranumerary embryos was hlgher arnong vigorous, suffe r competition at the time of natural1y formed fruits than among those ob­ germination. tained by artificial crossing. ACKNOWLEDGMENTS Seed germination: The percentages of germi­ nation arnong seeds obtained by artificial cross­ We thank Ernesto Paterniani (Department of ing were as follows: E. nocturnum 76.80%; E Genetícs, Escola Superior de Agricultura "Luiz erectum, 89 .61 %with a mean value of 83.30%. de Queiroz", Piracicaba, USP) fo r al10wing use The percentage of germination by crossing E. of the plants. We also thank the Conselho Na­ nocturnum and E. erectu m was 90.67%. Analy­ cional de Desenvolvimiento Científico e Tec­ sis of variance of these results yielded H = 7 .02 , nologico, CNPq for financial assistance. non significant, showing that the seeds result­ ing from the different crosses did not differ in germinating ability. The seeds removed from RESUMEN natural1y formed fruits showt'd the following En condiciones de invernadero, Epidendru m germination percentages: E. erectum 91.42%; noctumum Jacq. produce muchos frutos me­ and E. nocturnu m, 26.5 1%, with a mean value diante autopolinización y cleistogarnia. Esos of 58.96%. Thus, the perc:entages of germination procesos deben ser opcionales para el grupo, fo r seeds obtained by natural process (self-polli­ pues las polinizaciones intra- e ínter específicas nation or cleistogarny) were lower than those también conducen a la fo rmación de frutos que for seeds formed after intra- and interspecific contienen semillas con embrión. Las semillas artificial crossing (U=8 p<0,05 ).
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