The International Journal of Plant Reproductive Biology
(Indexed by CABI)
ISSN Print : 0975-4296; ISSN Online : 2249-7390
Volume-9, Number-2, Jan, 2017

Studies on the Gametophyte Development and Reproductive Behavior ofAcrostichum aureum L.

Meenam Bhatia, Priti, P. L. Uniyal1 and P.V. Madhusoodanan*
Department of Botany, University of Delhi, Delhi-110007, India
*Malabar Botanical Garden and Institute for Plant Sciences, Kozhikode-673014, India

Corresponding author:
Received: 11.01.2017; Revised: 13.02.2017; Accepted and published online: 01.03.2017


Acrostichum aureum is a pioneer species of mangrove sites growing rapidly and colonizing in disturbed habitats especially in the wetland areas. Plants of this species are useful in the mangrove restoration. This paper presents the study on the gametophyte development of A. aureum by light microscopy and scanning electron microscopy, including spore germination, development of the gametophyte and reproductive behavior. Spore is tetrahedral, trilete, and laesura is tenuimarginate. Spore germination is of Vittaria type. Germinal filament is short and uniseriate and prothallial development is Ceratopteris type. Gametophytes attained two-dimensional stage in 7-10 days and spathulate stage in about 15-20 days and became asymmetric chordate in 50-60 days. Gametophytes became bisexual after 50 days of sowing. In isolated populations 25% gametophytes and in composite population 68% produced sporophytes indicating the predominance of intragametophytic selfing. The length of unisexual phase before the attainment of bisexuality is sufficient to facilitate intergametophytic mating. It is further supported from observations on high numbers of sporophyte production and time of appearance of sporophyte in isolate and composite cultures. Gametophyte morphology and gametangial ontogeny also suggest that intergametophytic mating should be prevalent in gametophytepopulations of A. aureum.

Keywords: Mangrove, out-breeding, saline fern, restoration, trilete, tenuimarginate.

Volume : 9(2) pp. 88- 92, 2017 Download PDF

  Somatic embryogenesis in culture in vitro of Larix sibirica
Iraida Tretyakova*, Alena Ivanitskaya and Maria Park*
Laboratory of Forest Genetics and Tree Breeding, V.N. Sukachev Institute of Forest, Siberian Branch
Russian Academy of Sciences, 660036, Akademgorodok, 50, bld. 28, Krasnoyarsk, Russia

*e-mail :
Received : 28.04.2017; Accepted : 08.05.2017; Published online : 01.06.2017


Immature isolated zygotic embryos of Larix sibirica were experimentally cultured on AI medium (patent) containing L-glutamine, casein hydrolysate, ascorbic acid, and hormones (2.4-Dichlorophenoxyacetic acid and benzyladenine). The embryonal mass exhibited active proliferation during two-seven years on this medium with low concentration of cytokinins. The somatic embryos matured on the basal media with abscisic acid (120 mM) and polyethylene glycol. The embryological processes in L. sibirica consisted of elongation of somatic cells and their asymmetric division with formation of initial and tube cells. The embryo initial cells went under sequential divisions and formed small embryonal cells (embryonic globules). Suspensor-like structures developed from the edges of embryonic globules. Long-term proliferating cell lines and plantlets were obtained in L. sibirica. These lines produced from 2180 to 11103 somatic embryos (per 1g of callus fresh weight) during over two - seven proliferarion years. Somatic embryogenesis was strongly genetically controlled. Only free donor trees were becomes 23 embryogenic cell lines (Cl)

Keywords : embryogenic cell lines, somatic embryogenesis, Larix sibirica, proliferation, maturation, germination  
Volume : 9(2) pp. 93-100, 2017 Download PDF

Reproductive processes in Cembrae group pines on the clone plantation in
Krasnoyarsk forest-steppe

Galina V. Kuznetsova and Elena N. Muratova
V.N. Sukachev Institute of Forest, Russian Academy of Sciences, Siberian Branch. Krasnoyarsk, Russia
e-mail :;
Received : 10-05-2017; Accepted & Published online : 01.06.2017


The character of individual and geographical variability of growth, reproductive processes and seed producing of grafted trees of Cembrae group pines in a collection of clones established in Krasnoyarsk forest-steppe presented here. These are based on investigation on reproduction of clones of Cembrae group pines carried out for many years. Genetic determination of macro- and microstrobili characteristics in vegetative progenies has been shown. Yielded clones of Siberian pine were selected. Plantation of their seed progenies was established. Over-year seed producing dynamics at the plantation as a whole as well as dynamics of each provenance in specific growing conditions has been shown.

Keywords : Siberian pine, European pine, Scots pine, reproductive process, clone plantation, seed grafting plantation,
forest-steppe, graft, stock, microsporophyll, microstrobile

Volume : 9(2) pp. 101-108, 2017 Download PDF

Studies on pollen biology and stigma receptivity of Garcinia imberti Bourd.
(Clusiaceae)- a critically endangered tree of Western Ghats, Kerala

Rajkumar K.1, Keshavanarayan P.1, Shubharani R.1, 2 and Sivaram V.1, 2*
Laboratory of Apiculture and Biodiversity Conservation,

Department of Botany, Bangalore University, Bangalore-560056, India.
V Sivaram Research Foundation, No. 132, 2
nd Main, R.R. Nagar, Bangalore -560056, India.

Received: 14.03.2017; Revised : 01.05.2017; Accepted & Published online : 01.06.2017

Garcinia imberti Bourd. (Clusiaceae) is a critically endangered tree endemic to Agasthyamalai hills of Western Ghats, Kerala, India. In the present investigation, the pollen production, stigma receptivity, pollen viability, in-vitro pollen germination and pollen tube growth was studied during 2012 and 2014. The male flowers produced 7094 pollen grains per anther and 1, 41,890per flower.The pollen grains were round in shape with an average diameter of 20-36 μm. The stigma became receptive only after the opening of flowers. The pollen viability was 89.19±2.11% as tested by2% TTC. The in vitro pollen germination was 72.73±9.20% with 67.13±21.33% long pollen tubes in 20% sucrose solution. Higher pollen-ovule ratio (70945:1) and fruit and seed-seet only by cross pollination in self-incompatible Garcinica imberti indicated obligate xenogamy.

Keywords :Garcinia imberti, Clusiaceae, stigma receptivity, pollen production, viability, pollen germination, pollen tube

Volume : 9(2) pp. 109-114, 2017 Download PDF

Floral contrivances, pollination and breeding system of Clerodendrum inerme (L.)
Gaertn. (Lamiaceae)

Department of Botany, B.S.A. College, Mathura-281401, India

Received: 01.01. 2017; Revised: 12. 04.2017; Accepted and published online: 01.05.2017


Floral biology, floral contrivances, pollination ecology in Clerodendrum inerme (L.) Gaertn was studied. It is an erect to scandent or trailing evergreen shrub. Flowering commenced in the month of June and continued till October with optimum flowering in August, moderate in September and minimum in October. Flowers were bisexual, arranged in axillary cymes. Corolla 5, tubular and gamopetalous. Stamens 4 epipetalous, exserted and curved above. Pistil bicarpellary and syncarpous with inferior ovary, long style and bifid and wet stigma with unicellular smooth papille. Flowers open after 18:00 h and being protandrous the anthers dehisced soon after opening of flowers. Stigma became receptive on next day in the morning between 03:00–06:00 h and receptivity lasted till the afternoon on third day. Simoultaneously with the receptivity of stigma, a large quantity of nectar was secreted from peltate scales and unicellular trichomes present on the inner surface of calyx, corolla and ovarian surface attracting bees and butterflies for nectar or pollen or both. There were 3±1 ovules/ovary, anatropous with marginal placentation. Fruit a pear shaped drupe started developing in the month of July and lasted up to December. Flowers are protandrous and open in the evening followed by anthers dehiscence. On day 2, large quantity of nectar was secreted from peltate scales and unicellular trichomes present on the inner surface of calyx, corolla and ovarian surface attracting bees and butterflies for nectar and pollen. The flowers exhibited some morphological contrivances that help to avoid self-pollination. The flowers were highly protandrous as the anthers dehisce soon after the opening of flowers, and the stamens and stigma on a long style were in close proximity. while the bifid stigma either remains closed or bend away from stamens. In some other flowers, the stamens and style maintained some distance between each other. In the third type of flowers, the stamens were fully extended while the stigma on curved style was away from the anther lobes and in the fourth type of flowers, the stamens were curled inward while, the style was elongated. Pollen grains were tricolporate and tricolpate, sub-porolate, circular, echinate, and arranged as monods. Non-glandular unicellular trichomes and peltate scales were present on inner surface of calyx and corolla as well as on ovarian surface. Nectar secretion started along with the opening of flower (155±25.7 μL/flower) and gradually increased and was highest (214.72±35.12 μL/flower) between 08:00-11:00 h on next day. However, on day 3, during 11:00-16:00 h the quantity of nectar declined and was lowest (25±2.5 μL/flower). The nectar is dominated by glucose and fructose. Flowers were self-compatible, dichogamous and herkogamous. Bees (Apis cerana, A. dorsata, bumble bees (Bumbus lapidarius) and butterflies (Danaus genutia, Neptis hylas papaja and Eurema hecabe) foraged for both nectar and pollen, while hawk-moth (Macroglossum sp.) was a nectar thief. There was 25% fruit set in open pollinated flowers, 25% in geitonogamy and 75% in xenogamy. The pollen-ovule ratio of 720: 1 indicated facultative xenogamy.


Keywords: Floral contrivances, self-compatible, protandrous, dichogamous, herogamous, sugar dominated nectar, hawk moth,
bees and butterflies.

Volume : 9(2) pp. 115-124, 2017 Download PDF
  Studies on phenology and breeding system in Viola canescens (Violaceae)

Neha Mann*, P. L. Uniyal and Suman Lakhanpaul
Department of Botany, University of Delhi, Delhi 110007, INDIA

*e- mail:
Received : 22.03.2017; Revised: 23.05.2017; Accepted: 25.05.2017; Published online: 01.06.2017


Viola canescens (Violaceae) a medicinal herb is widely distributed in the temperate forests of Uttarakhand (India). The combination of chasmogamous and cleistogamous flowers in the plants of this species exhibiting mixed mating strategy makes its reproductive biology more relevant. Chasmogamous flowers appeared during March–May, were violet-white, pedicellate, pentamerous, zygomorphic, hermaphrodite and hypogynous, while cleistogamous flowers were born on the same plant in late summer on short horizontal peduncles close to the soil. The floral nectaries present only on chasmogamous flowers were examined for their structure, and nectar production. Several insects were observed to visit flowers and ants were found on the entire surface of plant during fruiting period indicating their role in seed dispersal.

Keywords : Chasmogamous, Cleistogamous, reproductive strategy, Uttarakhand
Volume : 9(2) pp. 125-128, 2017 Download PDF

Phenology, pollination mechanism, breeding system, seed dispersal and germination
in Aquilaria malaccensis Lam. a threatened tropical evergreen forest tree of North East India

N. Venugopal* and Ester Jones Marbaniang**
Department of Botany, Centre for Advanced Studies in Botany, North Eastern Hill University, Shillong-793 022, India

e-mail : *; **
Received: 01. 04. 2017; Revised: 28.05.2017; Accepted and Published online : 01.06.2017



Aquilaria malaccensis Lam. of Thymelaeaceae is an economically important tree for the production of agar oil. The species has been placed on Appendix II of CITES and belongs to the “Threatened” category of the IUCN Red List. Flowers are small and yellowish green and flowering takes place from March end to April first week followed by fruiting in May. Honey bees, beetles and thrips are the pollinators. The breeding system is xenogamous. In A. malaccensis wind and insects (ants) are the major dispersal agents of seeds. Seeds of A. malaccensis are recalcitrant and therefore, the percentage of viability, moisture content and germination of the seeds decreases with increasing the storage time, hence sowing of seeds soon after harvest is essential for better and higher percentage of germination. A. malaccensis is an evergreen tree but remain leafless from December to February due to caterpillars. The attack of caterpillars on the leaves and tender branches acts as a triggering mechanism for the formation of new leaves and buds.


Keywords : Aquilaria malaccensis Lam.; Thymelaeaceae; Threatened tree; Pollination; Breeding behaviour; Myrmecochorous
dispersal; Meghalaya; North East India.

Volume : 9(2) pp. 129-138, 2017 Download PDF
  Pollen Morphology and Physiology of Poa labillardieri (Poaceae)

Nabil M. Ahmad and Peter M. Martin*
Amenity Horticulture Research Unit, Plant Breeding Institute, The University of Sydney,
107 Cobbitty Road, Cobbitty, NSW 2570, Australia

Received : 24.04.2017; Revised: 22.05.2017; Accepted 25.05.2017; Published online:01.06.02017

Pollen grains of Poa labillardieri (Poaceae) were small, prolate, spheroidal and monoporate. The surface ornamentation consisted of spinulose islands separated by incised grooves. Average pollen size was about 24μm x 22μm in the hydrated state and was uniform in different genotypes. Optimum in vitro pollen germination in P. labillardieri was recorded in 25% sucrose solution supplemented with 10 mg/L H BO , 10 mg/L anhydrous CaCl , 10 mg/L KNO and 10 mg/L tetrasodium EDTA di-hydrate at pH 3 3 2 3 6.4. The methods used to assess pollen viability were compared with the optimum in vitro pollen germination in all the five genotypes. The results obtained with fluorochromatic reaction test (FCR) were more or less similar to the results of in vitro germination.
Keywords : Poa labillardieri, pollen morphology, pollen germination, pollen viability
Volume : 9(2) pp. 139-147, 2017 Download PDF
  Development of female gametophyte in Dioscorea caucasica (Dioscoreaceae)

 A. A. Torshilova, G. Yu. Vinogradova and G. E. Titova
Komarov Botanical Institute RAS, Saint-Petersburg 197376, Russia

Received: 05.05. 2017; Revised: 30.05.2017; Accepted and Published online: 01.06.2017


Present paper deals with the development of female gametophyte in Dioscorea caucasica (Dioscoreaceae). It has been established the tetrasporic Drusa-type, while earlier it was reported to be of monosporic Polygonum type. The characteristics of all the stages of embryo sac development up to mature embryo sac formation are described.

Keywords : Dioscorea caucasica, Dioscoreaceae, megasporogenesis, Drusa-type of the embryo sac development. Volume : 9(2) pp. 148-152, 2017 Download PDF
  Is sex irreplaceable? Towards the molecular regulation of apomixis

Vladimir Brukhin1, 2
Dobzhansky Center for Genome Bioinformatics, St. Petersburg State University 41 Sredniy Prospekt,
Vasilievsky Island, Saint Petersburg 199004, Russia

Department of Plant Embryology, Komarov Botanical Institute RAS, Prof.
Popov str. 2, Saint Petersburg 197376, Russia

Received: 03.05.2017; Revised: 15.05.2017; Accepted & Published online: 01.06.2017


Apomixis, defined as the asexual plant reproduction through seeds that results in the production of genetically uniform progeny and a natural way of cloning. Currently there are more than 400 plant species known to use apomixis as a strategy for their propagation. The primary fundamental aspects of apomixis are the bypassing of meiosis and parthenogenetic development of the embryo without fertilization. Apomixis attracts special attention because of its potential value for agriculture, as it could be harnessed for plant breeding programs enabling the permanent fixation of heterosis in crop plants. A better understanding of the molecular and genetic regulation of apomixis is important for developmental and evolutionary perspectives but also for implementation of engineering of apomixis traits into agricultural crop plants. Despite apomixis is considered as one of the key technologies for the improving agriculture, but currently how genetic and molecular regulation of this important trait occurs is not fully known. Recent information on the biology of apomixis and genes and genetic loci associated with the regulation of different components of apomixis is provided in the present review.

Keywords : apomixis, sexual reproduction, asexual reproduction, heterosis, fixation of hybrid vigour.
Volume : 9(2) pp. 153-169, 2017 Download PDF

Enhanced accumulation of phenolic acids in cryptogein co-transformed root
cultures of Digitalis purpurea L.

Amrita Basu and Sumita Jha*
Department of Botany, Calcutta University; 35, Ballygunge Circular Road; Kolkata 700019, India

Received : 29.05.2017; Accepted: 15.06.2017; Published online: 01.07.2017



The present work describes the effects of expressing a β-cryptogein gene on morphology and phenolic acids in hairy root cultures of Digitalis purpurea following Agrobacterium rhizogenes mediated transformation. The Ri-transformed and Ri crypt cotransformed root lines of D. purpurea showed rapid, ageotropic growth on solid MS medium. Morphologically, the roots expressing crypt gene in addition to rol genes were different than those not expressing crypt gene. The lateral roots in Ri crypt cotransformed roots did not elongate and the roots grew as entangled circular mass while the root lines obtained following transformation with wild type strain of A. rhizogenes showed elongation of laterals. The biomass accumulation of Ri crypt cotransformed root lines was higher than Ri-transformed root lines. The Ri-transformed and Ri crypt co-transformed root lines showed the presence of two phenolic acids viz. caffeic acid (CA) and ferulic acid (FA). The Ri crypt co-transformed root lines accumulated ~1.44 fold and ~2.5 fold higher CA and FA respectively as compared to the Ri-transformed root lines.
Keywords : Digitalis purpurea, cryptogein, transformed roots, phenolics
Volume : 9(2) pp. 170-174, 2017 Download PDF

Floral events related to the pollination of Hypenia reticulata
(Mart. ex Benth.) Harley (Lamiaceae)

Priscila Tunes¹ and Elza Guimarães²*
¹Graduation Program of Biological Sciences (Botany), São Paulo State University (Unesp),
Institute of Biosciences, 18618-970, Botucatu, SP, Brazil;
²*Department of Botany, São Paulo State University (Unesp), Institute of Biosciences, 18618-689,
Botucatu, SP, Brazil.*
Received :03.06.2017; Accepted & Published online : 01.07.2017



The Lamiaceae family presents 32 genus in Brazil. The Hypenia genus, Hyptidinae subfamily, comprises approximately 23 species that occur in seasonal and semi-arid regions in Brazil. The presence of flowers in which the stamens are encompassed by an articulated lower lip, can favour an explosive pollination mechanism. However, species of a same genus in this family can present different pollination mechanisms. Considering that floral morphology and functioning are closely related to species pollination, we aimed to describe the sequence of floral events related to the pollination of Hypenia reticulata such as (i) the mechanism of pollen and stigma exposure; (ii) the functioning of the lower lip; (iii) the process of style elongation; and (iv) the male and female functions during flowers anthesis. We conducted this study in a population of H. reticulata in a campo rupestre remnant in Botucatu, São Paulo state, Brazil, during two consecutive flowering seasons (2015-2016). For that, we isolated plants from the action of animals and wind; observed the flowers every hour from the beginning of anthesis until their senescence; determined the maximum length of styles and the time it took to achieve it; evaluated if style elongation was related to the release of the lower lip; performed stigma receptivity tests; and performed focal and direct observations to access which animal groups interacted with the flowers. Here, we described H. reticulata flower opening dynamics. We observed that H. reticulate is protrandrous, presenting an explosive pollination mechanism and temporal herkogamy. Our findings highlight the peculiarities of H. reticulata floral biology, mainly the mechanism of lower lip release, which revealed that its small protrandrous flowers rely on large- and small-sized biotic vectors for cross-pollination.


Keywords :Explosive pollination, floral biology, Hyptidinae, pollination ecology, protandry, temporal herkogamy
Volume : 9(2) pp. 175-178, 2017 Download PDF

(Short communication)
Modification of sex expression in Benincasa hispida Cong. by ethephone
(2-chloroethyle phosphonic acid) treatment

R. R. Singh
Department of Botany
, R.B.S. College, Agra-282002, India
Received :01. 02.2017; Revised: 01.03.2017: Accepted and Published online: 01.05.2017


Effect of foliar applications of aqueous solutions of ethepone (2-chloroethyle phosphonic acid) on sex expression in Benincasa hispida was studied. The seedlings at two and four leaf stage sprayed seven times with different concentrations of ethephone exhibited significant increase in the number of days taken for first staminate, and reduced days taken for pistilate flowering, the node position of staminate flowers was enhanced significantly but the reduction in the node position of first pistilate flower insignificant. The number of staminate flowers was reduced, number of pistilate flowers was enhanced significantly and the sexratio was significantly altered towards femaleness. The number of fruits/vine treated with lower concentrations of ethephone significantly increased, while the increase was insignificant in vines treated with higher concentration. Thus, ethephone is effective in enhancing femaleness in ash gourd.


Keywords : ethephone, staminate, pistilate, sex-ratio, node position.

  Volume : 9(2) pp. 179-182, 2017 Download PDF

  (Short communication)
On ecological aspects of the evolutionary reorganizations of Volvox ontogeny
A. G. Desnitskiy
Department of Embryology, Saint-Petersburg State University, Saint-Petersburg, 199034,
Universitetskaya nab. 7/9, Russia

Received: 16.05.2017; Accepted: 30.05.2017; Published online: 01.06.2017

Our previous investigations have shown that the green freshwater algal genus Volvox is characterized by considerable diversity in the details of asexual reproduction with respect to the rate, diurnal rhythms and light/dark control of asexual reproductive cell divisions. These traits, being simultaneously developmental and ecological, correlate with the data on latitudinal distribution of Volvox species. In an attempt to understand how this diversity might have arisen during evolution, we use the published data on paleoclimatology and continental drift in the past as well as some information on the migrating waterfowl, which are today the vectors responsible for green freshwater algae transportation and dispersal.

Keywords : algal geography, Cenozoic, eco-evo-devo, reproductive cell division, zygotes, Volvox.

Volume : 9(2) pp. 183-186, 2017 Download PDF


(Short communication)
Gemmae in Conocephallum japonicum (Thunb.) Grolle (Conocephalceae) and
Steph. (Anthocerotaceae) from Nagaland, North-East India


S.K. Chaturvdi
Department of Botany, Nagaland University, Lumami- 798 627, Nagaland, India

Received: 18. 05.2017; Revised: 12. 06. 2017; Accepted and Published online: 01.07.2017


In the present communication, multicellular discoid gemmae on Conocephalum japonicum (a thalloid liverwort) are reported. The gemmae are produced on the margins as well as on the dorsal surface of the thallus. On the other hand, stalked multicellular, discoid gemmae have been reported in Anthoceros angustus (Anthocerotophyta). They are produced on the margins as well as on the sub-marginal dorsal surface of the thalli.

Volume : 9(2) pp. 187-189, 2017 Download PDF
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