Clonal Propagation of Flacourtia indica for Ensuring Quality Planting Materials and Sustainable Supply of Edible Fruits

The present study was carried out at the Agriculture research field, Patuakhali Science And Technology University(PSTU), Patuakhali, from March, 2015 to April, 2016 to explore the domestication potential and to evaluate the rooting performance of Flacourtia indica (katabohori), a wild fruit species in Bangladesh, through clonal propagation by stem cutting under 3 different doses of rooting hormone IBA (Indole Buetaric Acid) and planted in the perforated plastic tray filled with coarse sand and gravel placed in the non-mist propagator.


Introduction
People of Bangladesh are generally poorly nourished despite substantial increase of food production in the country over past few decades.Most people suffer from malnutrition and resultant diseases.Ceaseless effort is therefore needed to improve the nutritional status and to increase food security, particularly for the rural poor (FAO, 1992).
Wild fruit trees offer vital insurance against malnutrition or famine during the seasonal food shortage and/or emergencies such as cyclone, floods, drought etc. though forest originated wild fruits are not consumed in greater quantities compared to main food staples.Wild fruits add variations in diets improve the palatability of staple foods and provide essential vitamins, minerals, proteins, carbohydrates and fats (Hossain et al., 2011).
Although the exact figure of the wild fruit species in the country is not known, Das (1987) compiled a list of sixty wild ed-ible fruit species from the forest areas of Bangladesh.Flacourtia indica (Burm.f.) Merr.commonly known as 'Baichi' or 'Katai', Synonymous to Flacourtia ramontchi L'Herit.(Family-Flacourtiaceae) is one of the important endangered fruit and medicinal plant species of Bangladesh.This is a bushy shrub or tree (Fig. 1) with a spiny trunk and branches.In shrub form it grows up to 25 feet (7.6 m) and as a tree it reaches a maximum height around 50 feet (15m).
The drooping branches bear oval leaves.Fruit is a small, red, fleshy, round berry.It turns a dark reddish-black when mature, and contains 4 to 10 brown, flattened, wrinkled seeds.The pulp is yellow or white and sweet with an acidic tang.It is eaten raw or made in to jelly or jam.It can be fermented to make wine.F. indica fruits are contained substantial amount of glucose and fructose as the main components.The main constitute of the polysaccharides in all the fruits was glucose and dietary fibre contents of the dry The experiment was laid out following a Randomized Complete Block Design (RCBD) with 4 treatments and 4 replications (blocks).The treatments were T 0 = control, T 1 = 0.2% IBA, T 2 = 0.4% IBA, T 3 = 0.8% IBA.The rooting ability of cuttings was significantly influenced by the application of IBA.The results showed that the highest rooting percentage (100) and maximum root number (8) of Flacourtia indica stem cuttings were obtained from the cuttings treated with 0.4% IBA followed by 0.2% IBA where as the longest root length (8.998 cm) was recorded with 0.2% IBA followed by 0.4% IBA.Survival percentage of the cutlings (the rooted cuttings) after 3 months of transferring them into poly bags was also significantly enhanced by exogenous rooting hormone (IBA) application.The highest survival percentage (84.5 %) was for the cuttings treated with 0.4% IBA followed by 0.2% IBA (71%).The similar trend was also observed for average cutling height and number of leaf.Findings of the present study reveal that the plant species is highly amenable for clonal propagation by stem cuttings using low-cost non-mist propagator.Considering both rooting percentage and root number, 0.4% IBA treatment of stem cuttings may be recommended for mass production of quality planting stocks for the domestication of the species in homestead agroforestry or in fruit orchards to provide edible fruit to rural poor people of natural disaster vulnerable Bangladesh.
Keywords: Flacourtia indica; IBA; clonal propagation; Non-mist propagator fruit was 29 to 79% but that fruit has by far the best combination of low free sugars and high dietary fibre (Nahar et al., 1990).
Flacourtia indica is an indigenous medicinal plant of Bangladesh (Figure 1) and India and it has been reported as an answer for the treatment of a variety of diseases and functional disorder.Fruits are used as appetizing, diuretic, and digestive, in jaundice and enlarged spleen.Barks are used for the treatment of intermittent fever also believed to be effective for arthritis.Roots are used in nephritic colic and gum is used in cholera (Kirtikar andBasu, 1998, Nazneen et al., 2002).The leaves and roots are used in herbal medicine for treatment of snakebite.
Most parts of the plant are used for cough, pneumonia, and bacterial throat infection.
It has also been used for diarrhea.There are different methods of domestication of plant species including various stages of development as mentioned by Booth and Turnball (1994).Among the stages, regeneration or propagation of the species play vital roles in the domestication process.Plants can be regenerated through seed germination or vegetative means like budding, grafting, stem cuttings or tissue culture.However, very little is known about the regeneration of the F. indica.The seeds are dispersed by birds.
To a limited extent the species is propagated from coppice.The species can be propagated from seed but little is known about germination techniques.Moreover, the information about the artificial regeneration of the spe-

Study location and climate
The study was conducted from at the

Agriculture research field of Patuakhali
Science and Technology University campus (lies between 21º48' and 22º36' north latitudes and between 90º08' and 90º41' east longitudes) Bangladesh (Figure 2) which enjoys typical tropical climate characterized by hot humid summer and cool dry winter.
The average temperature ranged between 14.3° to 27.4°C in winter and between 24.3° to 33.6°C during summer from 2010 to 2014 (Figure 3).The average annual rainfall was 2657 mm and varied from 1877 mm to 3120 mm (Figure 3) and rainfall usually takes place between June and September.The day length varies from 10 h 45 min in December to 13 h 25 min in June.Relative humidity is minimum (64%) in February and maximum (95%) in June to September.

Preparation of stem cuttings and setting up the experiment
The study on vegetative propagation was carried out in a low cost non-mist propagator (Figure 3).The non-mist propagator was constructed following the design described by Leakey et al. (1990) modified by Kamaluddin (1996)

Data analysis
Mean values for root number, root

Rooting ability
Rooting percentage of F. indica cuttings was significantly enhanced by exogenous rooting hormone (IBA) (Figure 4

Root number
Root number of cuttings was significantly enhanced due to the effect of applied IBA (Figure 6).The highest number of root (7.50) was developed in 0.4% IBA cies through vegetative propagation by stem cutting is very scarce and therefore extensive research in this aspect is needed for domestication considering its role in rural livelihood of Bangladesh.So, present study attempted to investigate the rooting ability of small leafy branch cuttings in low cost farmers' friendly non-mist propagator with or without rooting hormone IBA.

Figure. 1 .
Figure. 1. Full grown tree (a), 2 years old seedling at germplasm centre and ripe fruits of F. indica and was covered with sheet of transparent polythene such that the base was completely watertight and the lid was also airtight.The polythene base of the propagator was covered with a 10 cm thick layer of moist coarse sand mixed with successive layers of fine gravels and small stones.This layer supported rooting media kept in perforated plastic trays.Juvenile shoots of F. indica were collected from 2-years old stock plants raised in the Germplasm Centre of PSTU to get quality propagules.Two-node stem cuttings were prepared keeping two leaves intact without any trimming (considering smaller leaf area of F. indica) and then immersed briefly in a solution of fungicide Diathane M45 (Rohm and Co Ltd, France; 2 g/L in water) to avoid fungal infection.The cuttings were then rinsed and kept under shade for 10 minutes in open air for drying.As growth regulators are organic compounds that needed in the least amount which can support, inhibit, and may alter physiological processes of plants (Ulfa et al., 2013), stem cuttings were then treated with 0% (control), 0.2%, 0.4% and 0.8% IBA (Indole 3-Butyric Acid) solutions by dipping the cutting base into IBA solution to test effect of IBA on rooting ability and finally treated cuttings were planted into perforated plastic trays (12 cm depth) filled with coarse sand (Sylhet sand) mixed with fine

Figure 2 .
Figure 2. A geographical location of PSTU at Dumki Upazila
length and root diameter were calculated on experimental plot basis.All data were analyzed through computer based statistical program MSTAT-C (Michigan State University, East Lansing, MI, USA) following the basic principles, as outlined by Gomez and Gomez (1984).Significant effects of treatments were determined by analysis of variance (ANOVA) and treatment means were compared at 5% level of significance by Duncan's Multiple Range Test (DMRT).Microsoft EXCEL package (Microsoft Corporation, Pullman, WA, USA) were used for correlation and regression analysis.Rooting percentage was calculated and values were adjusted accordingly using arc sign transformation formula before putting the data into Analysis of Variance since the percentages of cuttings rooted were distributed between the range of 30 to 100 and proportions were based on equal denominator.

Figure 3 .
Figure 3. Non-mist wooden frame polythene propagator (a), Inside of propagator after setting of experiment (b), Plastic tray showing rooted cuttings(c), Rooted cuttings after transferring into growing media filled polyethylene bags treated cuttings and lowest (2.50) was in cuttings without IBA treatment.The findings of several authors supported the result of the present study that enhanced number of roots developed due to the IBA treatment.For instance, Hossain et al. (2002) mentioned that exogenous auxin (0.4% IBA) significantly (p 0.05) increased the root number of cutting of jackfruit.Abdullah et al. (2005) reported the highest root number in the cutting of Baccaurea sapida Muell.Arg.treated with 0.8% IBA solution.Dias et al. (1999) reported the highest number of root developed in the cutting of P. acerifolia treated with 6000ppm IBA 120 days after setting the cutting in the green house.The number of roots increased also with increasing IBA concentration for Neem (Azadirachta indica) cuttings treated with 0.2% or 0.4% IBA(Kamaluddin and Ali, 1996).AgainKamaluddin et al. (1996) recorded significant increases both in percentage rooting and number of roots with the application of IBA for Artocarpus heterophyllus.Again Kamaluddin et al. (1998) found that applied auxin significantly increased rooting ability of Chickrassia velutina cuttings.

Figure 4 .Figure 5 .
Figure 4. Rooting ability of cuttings of F. indica under various IBA treatments

Figure 6 .Figure 7 .Figure 8 .
Figure 6.Root numbers of F. indica cuttings developed under various IBA concentrations after six weeks from establishing the cuttings is one of the most important highly nutritious and valuable fruit species in Bangladesh that needs to be domesticated in homesteads for many reasons.Although propagation by seed is the prime method of plant multiplication, inadequacy of seeds as well as inadequacy of quality seeds is the major constraint that hampers plantation programs in the tropics.Flacourtia indica is also a mangrove forest originated wild fruit species of Bangladesh which never been cultivated in homesteads but were growing naturally.Due to increased population and shrinkage of coastal mangrove forest area the species has now reached almost an endangered situation.Due to scarcity of fruits or seeds vegetative propagation through stem cutting can be a better and helpful method of plant multiplication.The application of vegetative propagation in domestication of F. indica in homesteads of coastal areas of Bangladesh will ensure the production of uniform planting materials with desirable fruit characteristics (taste, seize, color), dis-

Table 1 .
Effect of IBA concentrations on survival, cutling height and leaf number of Flacourtia indica three months after transferring into polyethylene bags Figure 9. Growth performance of F. indica cutlings rooted due to IBA treatment three months after transferring into poly bags