or pollen, monoecious, candamarcensis, dioecious, Ambisexual, Key words: this speciesareproposed. management practices andpossible linesofresearchabout pollinators in orchards. Based on this information, some which ensures pollination without the need for male as plants, ambisexual in flowers male of occurrence Flowering offemaleplantscoincides withthepermanent 0.05). ≤ (P spring in 56% to decreases germination while summer, in 75% germinates plants, male and ambisexual of climatic conditions. Pollen from male flowers, from both the season; the female dioecious is maintained assuch,independently on depend to appears sexes different of flowers of existence plant. The ambisexual an as classified that are usually deformed. This condition allows them to be flowers bisexual as well as observed, are flowers male and female polygamous; are inflorescences plant Monoecious two during evaluated based on the factors of site, quantified season, and plant sex. and seasons. identified therefore were . Flowers frommonoecious and dioecious plants mountain of expression floral in variations the about learn to bedioeciousandmonoecious. The objective was to conditions inLaSerena(30°S,70° W), Chile,itisfound Ecuador, Colombia,andPeru.However, undercultivation is describedastrioeciousinthecentersoforigin (Vasconcellea pubescens A.DC.) ABSTRACT doi:10.4067/S0718-58392016000200001 Accepted: 16November2015. Received: 3July2015. * San JoaquínS/N,LaSerena,Chile. 1 María AngélicaSalvatierra-González ( ability inproductive mountain papaya Floral expression andpollengermination Corresponding author([email protected]). Instituto deInvestigaciones Agropecuarias, INIA Intihuasi,Colina Vasconcellea pubescens Vasconcellea pubescens. CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL In vitro pollen germination ability was also CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL Vasconcellea cundinamarcensis 1* , andConstanza Jana-Ayala - JUNE 2016

A.DC.)orchards and irrigationmanagement(Terra de Almeida etal.,2003). 1990), (Singh, humidity relative temperature, as such conditions other plants(AryalandMing, 2013),canbeaffected byclimatic like just expression, floral whose is notthecaseforC.papaya , This factors. climatic external with associated not are variations male flowers have a rudimentary pistil (Aguirre et al., 2009). These is attributedtothehypothesisofresourcecompetitionand because functional subdioeciousplants(Aguirreetal.,2007;2009). as defined are they flowers, perfect and male, female, have which mexicana (Muñoz, the of Other deformed 1988). produce that flowers bisexual scarce a as described dioecious (male andfemale plants) and monoeciousplantwith is and 2008), al., et (Carrasco plant trioecious a be to assumed is Chile in cultivated papaya Mountain flowers. of type other any report not did but plants, wild from flowers male and female recovered they Colombia; from V.flowers pubescens plants with polygamous inflorescences. Meza et al. (2011) describe for term inclusive plants, ambisexual and female both of consists that indicate (1972) Jiménez and Horovitz 2007). different types of flowers are described (Storey, 1941; Ming et al., 2012). related to the origin of the Andes Mountains (Carvalho and Rennes, that et al., 2007). The biogeographic history ofCaricaceae are Ecuador, Colombia, and Peru (Morales et al., 2004; Scheldeman male plantsbasedonuniquesequences(Gschwendetal.,2013). would have been separated into these different sequences for 2010); female and al., et (Wu sex determine that chromosomes the of Genetic studies ofVasconcellea havesuggestedtheevolution defines (2009) al. et 2013). Aguirre that it isderived fromadioecious ancestor (Aryal andMing, papaya, is C. papaya. The speciescultivated inChile,knownasmountain (V. monoecious monoica) andanotheristrioecious (V. is just like pubescensA.DC.) species one but (19), dioecious are which of and the most studied. The Vasconcellea 2012). and The INTRODUCTION Pistillate flowers of female and male also differ in size; this The centers with the greatest genetic diversity of Vasconcellea The trioecious sexual type is considered rare, and it is suggested Carica movesawayfromVasconcellea Knt t l, 05 Crhl ad Rennes, and Carvhalo 2005; al., et (Kyndt Vasconcellea plants have sexual polymorphism in which in polymorphism sexual have plants Carica papaya Carica papayaL.istheonlyspeciesofgenus A.DC., exhibit sexual variations in hermaphroditic trees, V. pubescens(Muñoz,1988;Scheldemanetal.,2007). - JUNE 2016 1 family consists of six genera, including genera, six of consists family Caricacea V. pubescensaspolygamous. family, such as and its diversification is has21species, most V. pubescens postulates Carica

136 RESEARCHRESEARCH inflorescence was carried out weekly on the sameplants the on out weekly axillary carried the developed was in of inflorescence flowers of Collection number inflorescence. greater each a collect to evaluation of inflorescence. axillary Monitoring occurredbetween January2012andJune2013. upper new a to moved was the basket finished, inflorescence selected the of flowering Once identified. was sex their and week per times three counted was flowers developed of number total flowers. The aborted of theseplantsunderwhichabasketwasinstalled to collect the evaluationfollowingyear. to start in order 2011 in August flowering first at identified were selected for each sex found in the orchards. Plants were these twolocations,threeplantswithsimilardevelopment At period. monitoring the during 65% of humidity relative mean and °C 16.5 of temperature mean a with similar were W), La Serena, Chile. Climatic conditions at both locations S; (29°56’34.8” Algarrobito 71°9’17.72” W) and Altovalsol is, (29°55’58.47” S; 71°7’9.89” that production, papaya orchards located inareas typically associated with mountain The studywasconductedintwoproductive mountain papaya MATERIALS ANDMETHODS appropriate fortheareaunderstudy. enable proposalsofagronomicrecommendations that are will knowledge plants. This male and ambisexual both from pollen germination ability was also evaluated in male flowers and female, their sex and number was identified in eachexpression of record; mountain papaya plants, defined as ambisexual of expression plant materialcultivatedinChileisnotconsidered. floral real if practices agronomic erroneous sexual type described in its centers of origin. This can generate to relying on types of sex associated with orchards in Chile during the seasons of the year, which leads date, To papaya mountain respectively. in expression floral of 1:1, reports no are there and 6:1 was ratio ambisexual nearby orchards(Altovalsoland Algarrobito), thefemale: two in Chile, Serena, La in example, For and orchards. papaya mountain of female in productivity affects which number plants, indefinite ambisexual an of consists orchard affects managementcostsevenmore.Finally, thecommercial simultaneously,expressed and not continues are practice this orchard atthebeginningofspring.However, becauseplants farmer thins outs the male plants the 8-10 mo after and planting in the sex unknown of seeds from originating plants four papaya, thenurseryprovidesaunitconsistingofthreeor et al.,2011). To overcomethischaracteristicofmountain hermaphroditic plantsarepreferredforfruitshape(Tamaki in occur not does this ; quality higher final affect plants production andfemaleplantsarepreferredbecauseoftheir female and ambisexual Both sex. plant by determined variables are which fruits, the of quality and number the by influenced is production orchard commercial CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL A modification was made during the second season thesecond during made was modification A An inflorescence with closed flowers was selected in each floral the about learn to was objective the that Given because relevance gaining is polymorphism Sexual or to the C. papayaortothe where C. papayawhere - JUNE 2016 (29°58’33” S;71°08’33” W), Valle delElqui. in Arid Zones(CEAZA)networklocated in Gabriela Mistral meteorological station of the Center for Advanced Studies data for thestudyperiodwasprovidedbynearest 2013 andMarch2014. Temperature andrelative humidity in the laboratory. Monitoring occurred between September sex their to according counted then were flowers different the Subsequently, season. previous the in tagged were that tube/total grains×100(FirmageandDafni,2001). in vitro of percentage The (40X). lens microscope the of fields three in germinated of (emission ofpollentube) and ungerminated pollen was counted grains of number the which in extracted was aliquot then An lux. 3000 and °C 24 at chamber growth h a 5 in for incubated and saccharose 5% of mL 2 a with with tube test extracted was stage Pollen each plants. at male and ambisexual were collected of flowers was male (2006) five out; al. carried et Parés-Martínez by established The procedure sepals. curved slightly had flowers when locations both at collected was Pollen sex. plant the and flowering of period was thattheabilitytogerminatevariesaccording in Pollen germinationwasmonitoredasareliabletestofviability Pollen germination seasons oftheyear (Figures1and2).However, the monoecious the during maintained was flowers the of sex the and study, no under orchards the with in observed were plants flowering in alterations female (1988), Muñoz with accordance In Observed typesofplants andflowers RESULTS ANDDISCUSSION the USA) statisticalprogram. with 0.05) ≤ Virginia, Warrenton, (P Technologies, (Statpoint means Statgraphics the compare to applied comparisons multiple was of test Duncan’s 2005). and Abak, and percentage a (Bal analysis as statistical the before √(%/100) to transformed expressed were data Germination analyzed. was Location and Season, Sex, factors three the for acompletely randomized design. The interaction of ANOVAby analyzed was germination Pollen location. and date recording each for flowers bisexual and female, male, the studysites. of each for independently processed was plants ambisexual were data and zero therefore notconsidered. The information about female and practically were during recordings reduced winter; is inflorescences axillary new of number The 72. were sex each of plants for inflorescences counted each typeofplantwereaveraged byrecordingdate. Total from seasons second and first the both in data Inflorescence Data processing statistical and analysis The analysis wasastatistical description (mean ± SD)for (Parés-Martínez et al., 2004); the hypothesis hypothesis the 2004); al., et (Parés-Martínez C. papaya germination was defined as grains of pollen with pollen

137 Figure 2.Seasonalvariations inthesexualexpression offemaleplantinflorescences in Algarrobito, Chile(means±SD). Figure 1.Seasonalvariationsinthesexualexpression offemaleplantinflorescences in Altovalsol,Chile(means±SD). agrees with the ambisexual plant definition (Horovitz and (Horovitz Jiménez, 1972)thatimpliespolygamousinflorescences. definition plant ambisexual the with agrees totally and 4), and 3 (Figures (1988) Muñoz by description the with agrees partly This flowers. bisexual with rarely and flowers, female and male both with inflorescences has plant Figure 3.Polygamousinflorescence withthree typesofflowers. CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL - JUNE 2016 (Figure 5). This ultimatelydeterminesfruitshape. plant female the of flower female the than zone ovary the in narrower are and shape and size in differ flowers female the female flowers found in the inflorescence. On the other hand, of number the surpass amply they and 2009), al., et (Aguirre pistil and 2005) arudimentary Miranda, and ‘Maradol’(Gil have for description the with coincides always this 4); (Figure flowers Male 1988). have adeformed carpel thatgives it acurvedshape(Muñoz, generally flowers bisexual and flowers female from mostly observe male flowers in specific periods in spring (Figure 8). observe maleflowers inspecificperiodsspring (Figure8). are mainly manifested during summer. It is only possible to flowers bisexual and female and season, the throughout pistil inflorescences are Polygamous dominated7). byand male6 flowers(Figures sites withstudy both a rudimentary at season the throughout expression sexual their in changes exhibited Plants Floral evolutionofambisexual plants In the observed polygamous inflorescences, fruits are fruits inflorescences, polygamous observed the In C. papaya

138 iue . eae lwr o a eae lf) andambisexual (left) afemale (right) plant. of flowers Female 5. Figure Figure 4. Types ofcommonflowersinpolygamousinflorescence foundin orchards inLaSerena. Figure 6.Seasonalvariations inthesexualexpression ofambisexualplantinflorescences in Altovalsol,Chile(means±SD). CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL - JUNE 2016 in observed inflorescences to similar is area study the in plants ambisexual in observed inflorescences flower regulated byhormones(Wu etal.,2010). be also can expression Floral study. present the in detected of expression such astemperature and relative humidity, which modify the factors, climatic with linked be may This flowers. male of expression an only is there when spring in especially year, and Ming, 2013) and could explain the differences within the Aryal 2010; al., et Wu 1990; (Singh, factors environmental various by influenced is the plants in expression Floral explain plants. can seeds, between inflorescence per flowers of number from and variations originated plants, papaya J. mexicanahermaphroditic plants, which,byproducing Variations from polygamous inflorescences to only male only to inflorescences polygamous Variationsfrom In theorchardsunderstudy, thegenotype of themountain flowers (OECD, 2005); this cannot be cannot this 2005); (OECD, flowers Carica V. pubescens

139 plant sex, male flowers have the same characteristics and characteristics same the have flowers male sex, plant the sites and the similar climatic conditions. With regard to of proximity the to attributed be can This 2). and (Tables1 location the or flower donor the from originating sex plant Pollen germination depended on seasonality and noton Pollen germination that flowers male promote adequatepollination. of existence permanent the of because availability pollen viable ambisexual have would flowers female plants, of behavior observed the Under 2009). al., et only male flowers, develop the three types of flowers (Aguirre humidity (%). Figure 8. Seasonal variations in the sexual expression of inflorescences observed at both locations based on temperature and relative Figure 7.Seasonalvariationsinthesexualexpression ofambisexualplantinflorescences in Algarrobito, Chile(means±SD). CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL RH: Relativehumidity, Temp: temperature,G.Mistral:Gabriela Mistral. - JUNE 2016 plants wouldproduce enoughviable pollen, especially in geographic location. of dependent 98.8% to 47.9% between viability pollen has that found (2008) al. et Caetano dueto temperature. ismainly and capacity germination influences that al. (2011), whoindicate that theseasonofyearisafactor Solo’‘Sunrise Tamakito hermaphroditic according the to et study. The degree of germination during summer is similar al., present the in et observed were flowers male perfect no 2006); (Parés-Martínez maleness perfect approaches type flower as increases germination pollen ‘Cartagena’, papaya produce pollen with the same germination capacity. InC. Finally, the proportion of male flowers in the ambisexual the in flowers male of Finally,proportion the V. pubescens

140 plants ofthemountainpapaya cultivated in Chile,aswell ambisexual of expression genetic the study to important be excluded intheproductiveorchards inthestudyarea. spring. Both situations would imply that male plants can be percentage of pollengermination ishigherinsummerthan the samegermination as thoseofmaleplants,andthe first observed. are flowers female when summer of beginning the at eliminating done that be should so post-planting at (males) flowers, plants undesirable male have only of beginning can the spring, at especially plants, Ambisexual fruits. summer when theyare functional andgiverisetodeformed and spring in mainly observed be can that flowers bisexual by some environmental variables. There is alownumberof influenced be could which year, of time on depending varied expression Floral flowers. bisexual and male, female, is, that inflorescence, polygamous a has plant monoecious dioecious (females and males) and monoecious plants. The In the study area, CONCLUSIONS male plantsunnecessaryforpollinationofthisspecies. of presence the make that flowers female of number greater a is there when is this temperature; higher with period the Season ×Sex Plant sex Season al 2 Pretg o ple griain n ae flowers male in germination pollen collected inthespringandsummer attwolocations. of Percentage 2. Table locations. ambisexual and male plants collected during two seasons at two fromflowers Tablemale of germination pollen Percentageof 1. Season ×Location Location Season P: probability, F:F-statistic,CV: to coefficientofvariation. according different significantly are Duncan’s test(P ≤0.05). columns the in letters Different P: probability, F:F-statistic,CV: to coefficientofvariation. according different significantly are Duncan’s test(P ≤0.05). columns the in letters Different Factor Factor CHILEAN JOURNAL OF AGRICULTURAL RESEARCH 76(2)APRIL In-depth research about this species is required. It would It required. is species this about research In-depth shows plants ambisexual of flowers male the from Pollen CV % P >F P >F Male Ambisexual P >F Summer Spring exhibited Vasconcellea pubescens CV % P >F P >F Altovalsol Algarrobito P >F Summer Spring Algarrobito 64.85a 66.11a 75.32a 56.08b 8.31% 0.063 0.4845 0 % Pollengermination % Germination 67.56a 64.85a 75.38a 58.09b 7.79% 0.366 0.3207 0 - Altovalsol JUNE 2016 70.03a 66.89a 75.42a 59.74b 7.36% 0.15 0.1429 0 cause offloraldifferentiation reportedinambisexualplants. naturalization of thisspeciessubjected to endogamy is the and a comparative study betweenV. foundinChile pubescens as evaluatetheirpossibleepigenetic responses. Similarly, Firmage, D., and A. Dafni. 2001. Field tests for pollen viability; pollen for tests Field 2001. Dafni. A. and D., Firmage, Carvalho, F.A., andS.Rennes.2012. A dated phylogeny ofthe B. García, R. Muñoz, P. Perez-Diaz, J. Avila, P. B., Carrasco, Caetano, C.M., T. Lagos, C. Sandoval, C. Posada,yD. 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