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Volume 6, Issue 3, June Issue - 2018, Pages:606-612


Authors: MAH Nagalakshmi, Sahana Vishwanath, KT Prasanna
Abstract: Vegetative propagation of Streblus asper an important medicinal plant, was studied for efficient rooting using different concentrations of auxins and  non auxinic Boric acid. Semi hard wood cuttings of   0.9 - 1.2 cm diameter from mature plants were collected and treated for 24 hours in liquid formulations of 50 ppm, 100 ppm and 200 ppm concentrations of IBA, IAA, NAA and 50 ppm, 150 ppm, 250 ppm and 350 ppm of  Boric acid individually and in combination. Results of the study revealed significant effect (p<0.05) of IBA and B combination on rooting and survival percentage and found that  50 ppm  and 10 ppm  IBA with 250 ppm of Boric acid induced highest  root number and length and significant  steckling capacity with high number of shoots than the other treatments and control. The study explores the scope of clonal propagation of S. asper by mature stem cuttings thus providing material for conservation of a selected clone.
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1 Introduction

The genus Streblus is one of the fascinating members of the ‘Fig’ family (Moraceae). It comprises of 25 species and it is geographically distributed mostly in tropical and subtropical Asia. In India it is represented by 5 species, among these 4 species are recorded from North east India while one species Streblus asper is found in drier parts, from Rohilkund, eastwards and southwards to Travancore (Roy et al., 2013). It is commonly known as Siamese rough bush and tooth brush tree (Rastogi et al., 2006). It is a dioecious tree on which flowering occurred between February - April and fruiting between May - July. The male flowers have rounded heads and greenish yellow color while female flowers are solitary and greenish (Plate 1A). The ripen fruits are yellowish, sweet flavoured, 1-seeded berry (Singh et al., 2015). Almost every part of S. asper is therapeutically used for various curative properties in Indian traditional folk medicine (Afjalus et al., 2013). Verma et al. (2016) reported efficacy of stem bark constituents in curing convulsive and depressive disorders. It is a rich source of cardiac glycosides; Tripathy et al. (2014) isolated twenty cardiac glycosides from the root bark of tree. Glycosides isolated from the root bark possess promising microfilaricidal activity. The seeds are reported to be beneficial in epistaxis and diarrhea; roots in epilepsy and inflammation and stem bark against lymphoderma, chyluria and other manifestations of filariasis (Hashim & Devi, 2002). The twigs are chewed to make brushes for cleaning teeth and to cure pyorrhea (Sanjay, 2015). Taweechaisupaponng et al. (2014) demonstrated that the ethanol extract of S. asper possessed inhibitory effect on subgingival biofilm formation  and suggested a potential for developing  a natural oral hygiene product against oral infection in people. The one seeded yellowish ripe berries  attract avian fugivores which include  babuls and Myna (Aruna & Balasubramanian, 2014). S. asper, being an economic and ecological important species, the propagation of this species through seeds is posing problem as the fruiting season is highly seasonal and thus available for only short period. Besides, the ripen fruits are eaten by avian predators and makes the task of seed collection difficult. Moreover, propagation of the species by seeds is time consuming, troublesome, and hence expensive (Baul et al., 2009). Because of these limitations to seed propagation, vegetative propagation of S. asper could increase its utility. There are no known reports in the literature of clonal propagation of S. asper. So, the main objective of the present study was to determine whether vegetative propagation by semi hard wood shoot cuttings could be developed as a practical alternative to seed propagation and the experiments were conducted to determine the effect of various auxins and Boron concentrations individually and in combination for the initiation, growth of root formation and initial growth performance.

2 Materials and Methods

The experiment was conducted in green house at the Botanical garden of the University of Agriculture Science, Gandhi Krishi Vignana Kendra, Bengaluru, India between March and July, 2017 with mean temperature range of 19 - 350C and relative humidity ranging from 42-90%, depicting typically tropical hot humid climate with 12 hour day and night cycle.

2.1 Preparation of cuttings and hormone treatment

Semi hardwood stem cuttings of seven node length and 0.9 - 1.2 cm diameter were obtained from healthy mother tree growing in Indian Institute of Science, Bengaluru (Dhuria, 2007). Before stucking three node length in the sand bed, all the leaves of cuttings were removed to reduce transpiration and cuttings were exposed for 24 hrs to various concentration of Indole-3-butyric acid (IBA), Indole-3-acetic acid (IAA), Naphthalene-3-acetic acid (NAA) in 10 ppm, 50 ppm, 100 ppm and 200 ppm and 50 ppm, 150 ppm, 250 ppm and 350 ppm of Boric acid (B). The vertical orientation of the stem cuttings was maintained and spaced adequately to allow sunlight. In the second trial, IBA which was found effective amongst three auxins tested, was combined with effective concentration of B (250 ppm) as suggested by Ono et al. (1999). Control was maintained by dipping the cuttings in distilled water devoid of hormones for 24 hours. Contrary to classic application, cuttings were not exposed to short time high hormone doses, but they were kept in low concentration of hormones for 24 hours. Stock solution of plant hormone of 1000 ppm was prepared by dissolving 100 mg of plant hormones in 0.5 N NaOH and made upto 100 ml by distilled water and desired concentration of rooting hormone was made by dilution of the stock.

2.2 Rooting media and experimental design

The cuttings were planted in a bed consisting of coarse sand mixed with fine gravel for rooting as it was found to be the best choice for rooting with good drainage and sufficient porosity to allow good aeration (www.itto.int)(Plate 1B). Gehlot et al. (2014) stated that aeration in sand medium plays a very significant role in number of root initiation and as  well as on root elongation. Three replications with 10 cuttings per replicate were planted for each treatment and were watered twice daily. A completely randomized design was adopted to study the influence of different treatments on rooting ability of cuttings.

 2.3 Transfer of rooted cuttings

After 12 weeks of setting the experiment, following the method of Hossain et al. (2018) rooted cuttings treated with IBA and B were transferred into poly bags containing a mixture of  sand, soil, cocopeat and perlite  in the ratio of 2:1:1:1 to assess steckling capacity  and initial growth performance in the nursery condition.

2.4 Data collection and statistical analysis

A cutting was considered to have rooted if it possessed one or more roots measuring not less than 1 cm. Data collected at the end of the rooting period were number of rooted cuttings per treatment, mean number/length of roots per cutting. After a period of 60 days of transplantation in polybags, number of shoots and survival percentage were recorded. Data collected were analysed using Graph pad prism 7. ANOVA and Dunnet’s multiple range tests were done to compare means from experimental groups against a control group mean to see the difference in effect.

3 Results

3.1 Percentage of rooting

The rooting percentage of S. asper varied from 6.66% to 46.66% under different treatments. In control, rooting was 6.66% and 93.34% cuttings remained dormant. Individually significant effect on rooting is shown by IBA 50 ppm followed by IBA 10 ppm and 250 ppm B (Figure 1). IAA was found to be less effective followed by NAA which was totally ineffective at all concentrations tested. In combination with B (250 ppm), IBA 50 ppm was found extremely significant followed by IBA 10 (P<0.05) (Figure 2). Though IBA was effective in inducing root and shoot formation at 10 and 50 ppm, only rooting was induced at IBA 100 ppm  and 200 ppm  without any shoot induction.

3.2 Number of roots of cuttings

The mean root number of S. asper cuttings varied from 0.66 to 9 across the hormone, its concentration and its interaction with Boric acid (B). Root number was lowest in control (Plate 1C) and  IBA at 10 ppm and  50 ppm with 250 ppm B (Plate 1D) had significant effect  on root number at P<0.05 (Figure 2a).

3.3 Length of roots of cuttings

The mean root length of S. asper cutting was significant at 50 ppm IBA + Boric acid and 10 ppm IBA + Boric acid combination. The highest mean root length was 3.5 cm and the lowest was 0.3 cm with IBA 50 ppm +B and control respectively (Figure 2b).

3.4 Shoot number of cuttings

IBA at 50 ppm and 10 ppm with 250 ppm B showed  significant affect on   shoot number and it was followed by IBA 100 ppm +B  (Figure 3a) .

3.5 Steckling capacity of rooted cuttings

Survival percentage of S. asper cutting was very significantly affected at 10 ppm, 50 ppm  and 100 ppm  of IBA combined with 250 ppm0 B  (Plate 1E & 1F; Figure 3b).

4 Discussion

The usefulness of auxins as a rooting aid during vegetative propagation has been summarized by many researchers (Ahmed et al., 2003 Abdullah et al., 2006; Sanjose et al., 2012). In Moraceae, auxin treatments were carried out in order to facilitate the rooting at different concentrations of IBA and found effective in rooting  of Ficus  and Morus  species which include F. glomerata L. (Bhatt & Badoni, 1993), F. Hawaii (Siddiqui & Hussain, 2007), M. nigra (Kalyoncu et al., 2009)  F. racemosa, F. microcarpa  F.religiosa, F. benghalensis (Mathew et al.,  2011), M. alba (Singh et al., 2014) and F. carica (Kaur & Kaur, 2017). Abdullah et al. (2005) reported rooting in few timber trees viz., Sweitenia macrophylla, Platanus acerifolia, Artocarpus heterophylla by

mature stem cuttings by using IBA. IBA is found to be more advantageous than other auxins, being non toxic in a wide range of concentrations without easily getting degraded with longer permanence in the application site (Ono et al., 1999). The application of IBA on rooting percentage of semi hard wood cuttings of mature S. asper had similar effect resulting in significant number of  robust and lengthy roots. Biochemically, induction of root using IBA was found due to activation of polysaccharide hydrolysis resulting in the increase of content of physiologically active sugar providing materials and energy for meristematic tissue and later for root primordia and roots (Abdullah et al., 2005). Besides, the use of water as a liquid carrier of IBA might be advantageous in the present study as reported by Ezekiel (2010) who stated that the preparation of liquid formulation of IBA dissolved in water is more effective for rooting most plants than IBA dissolved in alcohol as high concentration of alcohol may dehydrate and become toxic and injure the basal stems.

The occurrence of meagre rooting in non treated cuttings in S. asper is in conformity with rooting response of leaf less cuttings of matured stock plants of Albizia zygia, Blighia welwitschii (Egbe et al., 2012). The decline in ease of adventitious root formation of mature tree cuttings is may be due to physiological age of the stock plants integrated with several progressive changes in morphological, anatomical and biochemical traits such as decreased sensitivity of aging tissue to rooting promoters and/or accumulation of inhibitory substances which inhibit rooting and decreased content of endogenous auxins as stated by Ezekiel (2010). Exogenous application of IBA was found to result in root forming process in cuttings of S. asper to a maximum of 26% and was further enhanced by the addition of B (250 ppm) to 46% as stated by Abdullah et al., (2005) that though initiation of root primordial requires auxin but for subsequent primordial development both auxin and non auxin components are needed. Boric acid have a direct control  on the carbohydrate movement by the formation of  ionizable complex boron- sacarose,  thus facilitating the quick carbohydrate transport through the membrane to the places where cellular development and elongation occur (Ono et al., 1999; Stefanini et al., 2004). According to Esau (1965), boron plays a primary role activating the meristematic cells during the endogenous formation of root from pericycle or the endoderm. The effectiveness of boron with low concentration of IBA was supported by Ono et al, (1999) who stated that with B, IBA behaved effectively at low concentration.

The contrasting effects of increasing IBA concentration inducing only roots without sprout induction may reflect differential effects of applied auxin on the process of root initiation and development as the different concentrations of IBA applied gives rise to varied rooting response in different species. The highest survival (60%) in S. asper cuttings was observed when treated with 50 ppm IBA+B followed by 10 ppm IBA+B (50%) as observed in Baccaurea sapida cuttings (Nath & Barooah, 1992). The ANOVA and  DMRT  at P<0.05 showed that all the rooting parameters which include rooting percent, root number  and root length per cutting and steckling capability with well developed shoot were dependant on lower concentration of  IBA and  the presence of B at 250 ppm.

Conclusion

The male and female trees of S. asper cannot be detected before flowering of plants which takes 4-5 years of planting. Multiplication through mature stem cuttings from identified male and female matured stock plant can be an important tool for resolving the problem of female tree identification. In the present study, rooting of S. asper semi hard wood cutting was successfully achieved by the application of IBA and B (50 ppm +250 ppm respectively) and resulted in 60% steckling capacity. This protocol may be used in nurseries for its easy and faster multiplication. However, further experimentation with regard to the type of cutting, time of cutting and field trials of the rooted cuttings of the species for large scale clonal multiplication could be an important aspect of future study. The production of vigorous and healthy stecklings by vegetative means would enhance forest management through enrichment planting especially for dioecious species where male and more female trees with poor recruitment rates from seeds are to be planted together.

Acknowledgement

The author would like to acknowledge University Grants commission for the financial assistance to this research work under the sanction no. F.15-1/2016-17/PDFWM-2015-17-KAR-34814 (SA–II) dated 11/04/2016. The staff of department of Forestry and Environmental Science and Mahatma Gandhi Botanical Garden, University of Agricultural Science, Bengaluru are acknowledged for their support during the research work.

Conflict of Interest

The authors declare that there is no conflict of interest regarding the publication of this research paper.

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