T REPRO N DU LA C The International Journal of Reproductive Biology 10(1) Jan., 2018, pp.84-89 P T I F V O E

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S H Phenological behaviour of Piper sylvaticum Roxb. (), a medicinally T useful species occurring in Eastern Himalayan region Wahengbam Deepanita Devi, Padma Raj Gajurel* and Parakkal Rethy Department of Forestry, North Eastern Regional Institute of Science and Technology Deemed University, Arunachal Pradesh-791109, . *e-mail: [email protected] Received: 16. 09. 2017; Revised: 20.10.207; Accepted and Published online: 01.01.2018 ABSTRACT Phenology and flowering behaviour of a medicinally important dioecious Piper species i.e., Piper sylvaticum Roxb. (Piperaceae) found in eastern Himalayan region was observed. Vegetative and reproductive phenophases of both the male and female were studied. Different parameters like leaf initiation, leaf opening, initiation of flowers, peak flowering period, number of flowers, fruit formation and maturity, etc. were recorded. It was found that the leaves and spikes development are simultaneous and mostly occur throughout the year in male plants but varies in female plants. Peak flowering time was observed in the month of February and March in male and female plants respectively. Fruiting was observed almost throughout the year. Fruits mature in 4- 5 months. Keywords : Eastern Himalayas, Arunachal Pradesh, Piper sylvaticum, Medicinal plant, Dioecious, Phenology.

The genus Piper Linnaeus (Piperaceae) is represented by flushing and shedding as well as reproductive events such as about 2000 species (Quijano-Abril et al. 2006) which are bud formation, flowering and production of fruits (Francis et found in the pantropical region. This genus has three major al. 2007). It also provides the idea on the effects of clades (Jaramillo and Manos 2001), representing three large environment and selective pressure on flowering and fruiting geographical regions: America (1300 spp.), Asia (600 spp.) behaviour (Zhang et al. 2006). Keeping the view of the and the South Pacific (100 spp.) (Quijano-Abril et al. 2006). importance of phonological studies in cultivation, The new world Piper species are bisexual and are trees or management and conservation of plants, a study have been bushy shrubs whereas, the old world Pipers are mostly undertaken to understand the different phenological events of dioecious and climbers. Over 100 species of Piper have been Piper sylvaticum considering both male and female plants reported from India, distributed in Western Ghats and Eastern occurring in the north east and East Himalayan region. Himalaya with about 65 species in north eastern states (Gajurel et al. 2008). The genus contributes economically and MATERIAL AND METHODS ecologically valuable species, and has been commercially used The present study was carried out at NERIST campus, as spices and medicines. Because of their economic potential Arunachal Pradesh that is located between 27°07’ N latitude, the species are preferred in management and conservation and 93°22’ E longitude and situated 126 m a.m.s.l. during the programs in tropical areas. Piper nigrum Linnaeus, P. betle years 2013 and 2014. Different phonological events of both Linnaeus, P. longum Linnaeus and P. methysticum G. Froster male and female plants of P. sylvaticum were carried out in the are the well-known economically important species, besides plants raised in the nursery of Forestry Department, NERIST many species of medicinal and ethnobotanical uses (Jaramillo which were collected from different parts of Arunachal and Manos 2001, Quijano-Abril et al. 2006, Gajurel et al. Pradesh. To observe different phenophases, 25 individual 2008). Many other wild species of Piper (P. mullesua plants each for male and female were tagged for the detail Buchanan- Hamilton ex D. Don, P. pedicellatum C. DC., P. study. Flowering and fruiting phenology were studied by beteloides C. D. C, P. wallichii (Miquel) Handel-Mazzetti, randomly tagging 30 spikes each from both the sexes. etc.) have been used as medicines for the treatment of common Continuous observations on leafing (include initiation, diseases or as preservative (Gajurel & Rethy 1998). The opening, peak period of leaf formation, time taken for species P. sylvaticum Roxb is a common wild Piper found maturity, size of mature lamina, number of new leaves extensively in north east India and the fruits are used as developed, senescence and fall); flowering (spikes formation, substitute for commercial Piplai (P. longum) in Ayurvedic flower initiation, maturity of spikes and flowers, peak formulation and also as in indigenous medicine (Kirtikar and flowering period, maturity of a flowers, length of the mature Basu 1933, Gajurel et al. 2008, Babu et al. 2016). spike, number of flowers in a spike, completion of flowering Phenology is generally described as the art of observing in a spike, duration of flowering in a spike and number of the phases of the life cycle or activities of organism as they flowers per spike) and fruiting (fruit initiation , fruit maturity, occur throughout the year (Leith 1973). For plants, recurrent dropping of fruiting spikes, etc) were made periodically for biological events include vegetative processes such as leaf one full calendar year for recording phenological data. The 2018 Phenological behaviour of Piper sylvaticum Roxb. (Piperaceae) 85

different phenological events were observed in regular cancer (Wang et al. 2014, Shahriar et al. 2014, Bose et al. intervals for each phenological event following standard 2015). The root of it is used as an antidote against snake poison methods (Marquis 1988, Kasarkar & Kulkarni 2011, Nanda et in the indigenous India (Kirtikar and Basu 1933). Other than al. 2014). All the recorded data were then analysed and medicinally important, the species are used as condiments and presented. For vegetative phenology the developmental stages vegetable (Srivastava and Adi community 2009). of leaves were observed only in the plagiotropic fertile shoots. Phenology—Detail observations on vegetative phenophases are presented in Table 1. It was observed that leaf RESULTS initiation in the species were started from the month of December and continued till May in male and June in female Habit—Piper sylvaticum Roxb. is a dioecious, creeping plants. The peak period of leaf initiation was found in the or low climbing shrub with dimorphic branches (Fig. 1) found month of January in male and February in female plants. On in North East region of India and other neighbouring countries average, about 9 to 10 days time was taken for opening of leaf like , Bhutan, Burma, Nepal and . Leaves in from the day of leaf initiation in both male and female plants. vegetative branches- membranous or thinly coriaceous, finely Number of new leaves developed per plant varies from 10 to puberulent broadly ovate- cordate or oblong, acuminate at 21with an average of 15.64 + 04.38 in male and 10 to 15 with apex, base deeply cordate with round lobed, petiole with a an average of 11.48 + 02.80 in female plants. New initiated deep groove; in fertile branches leaves are elliptic lanceolate, leaf opened after an average of 9 and 10 days from the day of shortly acuminate, petiole short. Spikes erect with densely leaf initiation in male and female plant respectively. It takes 3 arranged flowers, white or pale in colour; rachis hairy. Male to 4 months to obtain the full maturity size of a leaf. Leaf spikes long cylindrical or filiform with large number of male senescence starts from the month of November till March in flowers consisting 2-3 short stamens. Female spikes short male plant whereas in female plant, it starts from December cylindrical, consisting numerous globose ovary. Fruiting and continue till June. Leaf fall started in the month spikes short cylindric. Drupes densely aggregated, spherical. from September till April in male and October to May in Medicinally the species is used in the treatments of various female plants. Size of the matured leaf lamina was about diseases such as cough and cold, cut and wounds, laxative, 10.65 x 3.84 cm and 11.34 x 5.74 cm in male and female plants anthelmintic, carminative, bronchitis, spleen, tumors and respectively.

Table 1- Vegetative phenophases of male and female plants of Piper sylvaticum. Sl. No. Vegetative phenophases Male Female 1 Leaf initiation period December to May December to June 2 Peak period of leaf initiation January February 3 Time taken for leaf opening after initiation (days) 9.2 + 2.373 9.87 + 1.96 4 Time taken for leaf maturity (month) 3 to 4 3 to 4 5 Time taken for senescence of matured leaf (month) 1 to 2 1 to 2 6 Leaf fall period September to April October to May 7 Size of a matured leaf (cm) 10.65 (+ 0.80) x 3.84 (+ 0.38) 11.34 (+ 0.63) x 5.74 (+ 0.59) 8 Number of new leaves developed in the fertile branch 15.64 + 04.38 11.48 + 02.80

Flowering phenology—It was observed that the species shows initiation of spikes and 30 days in female plants. In male plants, seasonal variations in flowering of the male and female plants. anther dehisces after 13 days of flower initiation i.e., maturity of Developing stages of male and female flowering spike are flowers took around 42 days while in female plant it took only presented in Fig. 2 (a-j). In male plants, the flowering spikes 13 days. Flowering process was completed in 42 and 24 days in were observed almost throughout the year but in the female male and female spikes respectively after the 1st day of initiation plant, the flowering spikes were developed during the month of of flower in a spike. Time taken for elongation of spikes was 79 December to April. It was found that all the individuals bear and 53 days in male and female spikes respectively. Male spikes spikes in the month of February and March for both male and were fallen after 12 days of maturity. So, the overall life span of a female plants (Fig. 3). Highest numbers of budding spikes were spike is 92 and 53 days in male and female individuals recorded in the month of January in male and in February in respectively (Table 2). Number of spikes per plant ranged from female plants. The average number of new budding spikes 10 to 21 with an average of 13 in male plants and in female recorded in a lateral fertile branch in the month was 04.76+02.57 plants the number ranges from 10 to 15 spikes with an average of in male and 05.16+01.75 in female plants (Fig. 4). While during 11 in female plants. The average length of the male spike was the peak flowering period the average number of male flowering 6.39 cm with 664.77+ 218.56 flowers per spikes and 1935 spikes were recorded as 03.92+ 02.72 and 04.12+ 1.94 in female stamens per spikes. The average length of the female spike was plants in the month of February and March respectively (Fig. 5). 1.19 cm with 71.07 flowers. The female spikes do not fall and In male plants, flower initiation was observed after 37 days of develop into infructescences. 86 The International Journal of Plant Reproductive Biology 10(1) Jan. 2018, pp.84-89

Table 2. Flowering phenophases of male and female plants of Piper sylvaticum. Sl. No. Flowering phenophases Male Female 1. Flowering season Throughout the year December to April 2. Pick period of flowering February March 3. Time taken for flower initiation after initiation of spike (days) 37.27 + 17.72 29.60 + 13.65 4. Time taken for maturity of a flowers (days) 12.47 +7.34 12.27 + 3.63 5. Completion of flowering period in a spike from the 1st day of 42.00 + 19.25 23.60 + 6.10 initiation of flower in a spike (days) 6. Time taken for spike elongation (days) 79.26 + 21.85 53.20 + 19.75 7. Time taken for falling off of a matured spike (days) 12.27 + 7.87 NA 8. Number of spikes per plant 10 to 21 10 to15 9. Total life span of a flowering spike (days) 91.53 + 21.78 53.20 + 19.75 10. Length of the matured spike (cm) 6.39 + 2.33 1.19 + 0.40 11. Total number of flower / inflorescence 664.77 + 218.56 71.07 + 15.23 12. Number of anthers per spike 1935 NA

a b

Fig. 1- Piper sylvaticum (a) Male plants (b) Female plants

Fig.2– Different developmental stages of flowering and fruiting spikes in P. sylvaticum- Female spike (a-e): (a) initiation of budding (b) budding spike, (c) initiation of flower, (d) mature flowering spike (e) female flowers; Male spikes (f-j): (f) initiation of budding, (g) budding spike, (h) initiation of flower, i) mature spike (j) male flowers; Fruiting spike (k-n): (k) fruiting initiation in female spike, (l) completion of fruiting, (m) fruiting spike with developing fruits (n) fully matured fruiting spike. 2018 Phenological behaviour of Piper sylvaticum Roxb. (Piperaceae) 87

infructescences starts shortly after 9 days of ripening of fruits. The dropping off season for infructescences was recorded during the month of September to February. The overall fruiting period was 263.7 days i.e, around 9 month (Table 3).

Table 3. Fruiting phenophases of Piper sylvaticum.

Sl. Fruiting Phenophases No. 1 Fruiting season Throughout the year 2 Fruiting initiation period January - May 3 Peak fruiting initiation period April Fig. 3—Temporal variation of male and female plants of P. sylvaticum 4 Time taken to initiate fruit from the 1st 23.60 + 6.10 bearing spikes in different month. day of flower initiated (days) 5 Time taken for completion of fruiting in 35.20 + 7.97 a spike after initiation of fruiting (days) 6 Time taken for ripening of fruit from the 254.67 + 39.40 date of initiation of fruit (days) 7 Time taken for dropping off of matured 9.03 + 0.30 infructescences (days) 8 Infructescences dropping period September to February 9 Total fruiting duration (days) 263.69 + 8.78

Fig. 4— Number (mean value) of male and female budding spikes in different month.

Fig. 6– Temporal variation of the female plants bearing infructescence.

Fig.5—Number (mean value) of male and female flowering spikes in different month. Fruiting phenology—The female spikes turn into the fruiting spikes and were seen almost throughout the year but with different maturity stages. The details of developing stages of infructescences are shown in Fig.2 (k-n). It was observed that during the month of March to October, 100 % of the female plants bear infructescences (Fig. 6). The fruiting initiation Fig. 7– Fruits development period. started from January and continue upto May with peak fruiting initiation period in April (Fig. 7). It took around 24 days to DISCUSSION initiate fruit in the female flower from the day of flowering and a long fruiting duration time was observed. In a spike the The present study on the phenology of both the sexes of fruiting completed in 35 days after the day of fruit initiation P. sylvaticum revealed that the leaf flush in fertile branches start. For fruit maturity and ripening it took almost 255 days synchronised with flowering period and observed in a specific from the day of fruiting initiation. However, the dropping of period. The result is in contrast with the other bisexual Piper 88 The International Journal of Plant Reproductive Biology 10(1) Jan. 2018, pp.84-89

species of the new world (Opler et al. 1980, Marquis 1988, compared to female flowering spikes that ensure the pollen Thies and Kalko 2004, Valentin–Silva and Vieira 2015) and in supply during the flowering period of the female. Fruiting P. mullesua of the old world where leaf flush occur throughout spikes were found almost throughout the year which is the year (Devi et al. 2016). Leaf flush synchronization with significant for their utilization as the fruits are very commonly the flowering spike production, are marked as phonological used for their medicinal values and have commercial demand. characteristic in seasonal tropical forests (Thies and Kalko Acknowledgements—Authors are thankful to the Director, 2004). Production of more flowers in males than the female NERIST for the research and nursery facilities. The authors plants in dioecious species is a near universal pattern (Gracia would like to thank Mr. Dilip Deori for his help in the plantation and Antor 1995, Guitian 1995). In P. sylvaticum too, the same and management of Piper plants in the nursery and Mr. Kh pattern has been observed. Flowering period was found Ronald for his help in data analysis. different between the male and female individuals. In P. sylavticum male flowering period start earlier than the female REFERENCES flowering period. Fruiting starts in January and continues Anderson GJ, Bernardello G, Opel MR, Santos-Guerra A and throughout February-May. In some dioecious plants male Anderson M 2006. Reproductive biology of the dioecious flowering period start earlier than the female flowering period Canary Islands endemic Withania aristata (Solanaceae). (Bawa 1983). It has also been reported that the Piper species Am. J. Bot. 93(9) 1295 – 1305. can modify their phenological responses according to their distributional area. For instances fruiting season of P. Babu V, Singh SK, Venkatasubramanian P and Gajurel PR arboreum in Brazil (Bizerril and Raw 1997) and P. amalago in 2016. Comparative analysis of major alkaloids in Piper Costa Rica (Fleming 1981) were different from that of Santa species traded as ‘Pippali’ in South Indian markets: Genebra. Spike developments and flower formation were absence of the chief known constituent- piperine in observed throughout the year only in the male plants with one selected samples. Curr. Sci. 111(9) 1507 – 1510. peak period, same has also been observed in other new world Pipers like P. arieianum (Marquis 1988), P. cernuum (Mariot Bawa KS 1983. Patterns of flowering in tropical plants. In: et al. 2003) and P. vicosanum (Valentin-Silva and Vieira Jones CE and Little RJ (eds.), Handbook of Experimental 2015). In female plants the spike and flower formation were Pollination Biology. Van Nostrand, Reinhold, New York. observed only for five months a shorter period in comparison Pp. 394 – 410. to the male plants. The peak season of flowering of both the sexes of the studied species were observed in low rainy season Bizerril MXA and Raw A 1997. Feeding specialization of two i. e., in the month of February and March. Gajurel (2002) also bats and the fruit quality of Piper arboretum in Central recorded the similar flowering time in many Piper species of Brazilian gallery forest. Revista de Biologia Tropical the region. The Piper of central and South America also 45(2) 931-918. usually flowers in dry season (Opler et al. 1980, Fleming 1985, Marquis 1988, Marinho-Filho 1991, Wright 1991). Like Bose D, Roy JG, Mahapatra (Sarkar) SD, Datta T, Mahapatra, Piper, the other dioeceous plant like Withania aristata also SD and Biswas H 2015. Medicinal plants used by tribals flowers simultaneously on both sexes year round (Anderson et in Jalpaiguri district, West Bengal, India. J. Medicinal al. 2006). Occurrence of more than one flowering episode has Plants Studies, 3(3) 15-21. been observed in many other species like Trichilia pallida Devi WD, Gajurel P R and Rethy P 2016. Phenology of Piper (Morellato 2004), Ficus sp. (Jia et al. 2007), etc. The mature mullesua Buchanan-Hamilton ex D. Don (Piperaceae) - a male individuals produce more number of spikes as compared medicinally important dioecious plant. Pleione 10(2) to female individuals. The number of spike formation by a 239-247. plant depends on the age of the plant. However, the health of a plant may also become a determinative factor in flowering and Fleming TH 1981. Fecundity, fruiting pattern, and seed fruiting. dispersal of Piper amalago (Piperaceae), a bat-dispersed Infructescences in P. sylvaticum were observed round the tropical shrub. Oecologia 51(1) 42-46. year with a longer period of fruit maturity. Snow (1965), Thies and Kalko (2004) and Devi et al. (2016) also observed Francis QB, John P, Suriantata, Laszlo N, Gemma D and sequential fruiting periods in Piper species that relates to the Benedict CV, 2007. Reproductive phenology over a 10- year-long fruit production. In the Ficus species also the fig year period in a lowland evergreen rain forest of central crops bore several times throughout the year (Zhang et al. Borneo. J. Ecol. 95 828 – 839. 2006, Kuaraksa et al. 2012). From the result of the study, it is clear that the spike Gajurel PR 2002. Experimental taxonomic studies on Piper production is directly correlated with leaf production. It is also species of Arunachal Pradesh. Ph.D. thesis, North clear that male flowering spikes have long flowering season as Eastern Hill University, Shillong, India. 2018 Phenological behaviour of Piper sylvaticum Roxb. (Piperaceae) 89

Gajurel PR, Rethy P, Kumar Y and Singh B 2008. Piper Morellato LPC 2004. Phenology, sex ratio, and spatial species (Piperaceae) of North-East India (Arunachal distribution among dioecious species of Trichilia Pradesh), Bishen Singh & Mahendra Pal Singh, Dehra (Meliaceae). Plant Biol. 6 419 – 497. Dun. Nanda A, Suresh HS & Krishnamurthy YL 2014. Phenology Gajurel PR and Rethy P 1998. Piper species of Arunachal of a tropical dry deciduous forest of Bhadra wildlife Pradesh and its potential for development as a sanctuary, southern India. Ecol. Processes 3(1) 1-12. commercial crop in the state. Arunachal Forest News 16(1, 2) 32 – 37. Opler PA, Frankie GW and Baker HG 1980. Comparative phenological studies of treelet and shrub species in Garcia MB and Antor RJ 1995. Age and size structure in Tropical Wet and Dry Forests in the Lowlands of Costa populations of a long-lived dioecious geophytes: Rica. J. Ecol. 68(1) 167 – 188. Borderea pyrenaica (Dioscoreaceae). Int. J. Plant Sci. 156 236 – 243. Quijano-Abril MA, Callejas-Posada R and Miranda-Esquivel DR 2006. Areas of endemism and distribution patterns for Guitian P 1995. Reproductive biology of Rhamnus Neotropical Piper species (Piperaceae). J. Biogeog. 33(7) legionensis Rothm., a dioecious species endemic to the 1266 – 1278. northwest Iberian peninsula. Flora 109 1 – 8. Shahriar Md.H, Chowdhury ASMHK, Rahman Md.S, Jaramillo MA and Manos PS 2001. Phylogeny and patterns of Uddin Md.P, Al-Amin Md., Rahman Md.M, Bhuiyan floral diversity in the genus Piper (Piperaceae). Am. J. Md.TA, Afrin S, Chowdhury S, Masud-ur-Rahman, Bot. 88(4) 706 – 716. Md., Azad AK and Rahmatullah M 2014. Scientific validation of medicinal plants used by a folk medicinal Jia XC, Dan Y, Zhang Y and Chen YZ 2007. Direct evidence for the cycling of ûg wasps within one male ûg tree. practitioner of Chuadanga district, Bangladesh. Nordic J. Bot. 25(1-2) 119-124. World J. Pharmacy and Pharmaceutical Sci. 3(11) 13-24. Kasarkar AR and Kulkarni DK 2011. Phenological studies of family Zingiberaceae with special reference to Alpine and Snow DW 1965. A possible selective factor in the evolution of Zingiber from Kolhapur region (MS) India. Biosci. fruiting seasons in tropical forest. Oikos 15 274 – 281. Discov. 2(3) 322 – 327. Srivastava RC and Adi community 2009. Traditional Kirtikar KR and Basu BD 1933. Indian Medicinal Plants knowledge of Adi tribe of Arunachal Pradesh on plants. (2nd edn.), Vol. 3. Lalit Mohan Basu Publications, Indian J. Traditional Knowledge 8(2) 146-153. Allahabad, India. Thies W and Kalko EKV 2004. Phenology of neotropical Kuaraksa C, Elliott S and Hossaert-Mckey M 2012. The pepper plants (Piperaceae) and their association with their phenology of dioecious Ficus spp. tree species and its main dispersers, two short-tailed fruit bats, Carouia importance for forest restoration projects. For. Ecol. perspicillata and C. castanea (Phyllostomidae). Oikos Manag. 265 82 – 93. 104(2) 362 – 376.

Leith CE 1973. The standard error of time-average estimates Valentin-Silva A and Vieira MF 2015. Phenology of two co- of climatic means. J. Applied Meteorology and occurring Piper (Piperaceae) species in Brazil. Climatology 12 1066-1069. Australian J. Bot. 63(7) 581 – 589. Marinho-Filho JS 1991. The coexistence of two frugivorous Wang Y, Morris-Natschke SL, Yang J, Niu H, Long C and Lee bat species and the phenology of their food plants in K 2014. Anitcancer principles from medicinal Piper Brazil. J. Tropical Eco. 7(1) 59-67. plants. J. Traditional and Complementary Medicine 4(1) 8-16. Mariot A, Mantovani A and Reis MS 2003. Uso e conserva¸cão de Piper cernuum Vell. (Piperaceae) na Mata Atlântica: I. Wright SJ 1991. Seasonal drought and the phenology of Fenologia reprodutiva e dispersãode sementes. Revista understory shrubs in a tropical moist forest. Ecology Brasileira de Plantas Medicinais 5 1 – 10. 72(5) 1643-1657. Marquis RJ 1988. Phenological variation in the neotropical Zhang G, Song Q and Yang D 2006. Phenology of Ficus understory shrub Piper arieianum: causes and racemosa in Xishuangbanna, Southwest China. consequences. Ecology 69(5) 1552 – 1565. Biotropica 38(3) 334 – 341.