Bogor, 21-22 October 2015
774 Moreover, cytokinin and auxin are often required to achieve quantitative growth increase of
cell number and volume. Cytokinin benzylaminopurin BAP particularly stimulate protein synthesis and participate in cell cycle control. They can promote the maturation of chloroplast
and delay the senescence of detached leaves George de Klerk, 2008; Wróblewska, 2013. Auxin indol butiric acid IBA promotes, mainly in combination with cytokinins, the growth
of calli, cell suspensions and organs, and also regulate the direction of morphogenesis. They are involved in the formation of maristems giving rise to organs Machakova, 2008.
For many forest tree species, the level of juvenility is not demonstrated by propagules from trees of selectable age. Two alternative approaches to the problem are 1 rejuvenation, the
derivation of juvenile propagules from trees of selection age, followed by multiplication of the selections and 2 maintenance of juvenility for the period of a cloned test, followed by
multiplication of the best clones include long-term hedging Libby et al., 1972. For some trees, for example many species of Eucalyptus and Araucaria, the stimulation of the
development of coppice shoots from juvenile meristems which persist near the base of the stem is the simplest approach. Strategies incorporating the maintenance of juvenility are
appropriate then for most of the major industrial species. The ability to rejuvenate selections made on the basis of an index would undoubtedly confer added flexibility, but would only be
really beneficial where heritability was high, andor where clone testing was difficult. This might apply to selections for some stress tolerant trees, wood quality traits, and also as an
interim measure for long rotation species, while results of lengthy cloned tests are awaited Haine, 1994; Ballester et al., 1999; Basher Salima, 2007.
In the present study, vegetative rejuvenation formation of gall rust tolerant in an effort to sustain the availability of juvenility for explants, sprouting ability of branch and observation of
the initiation response of their rejuvenation axillary buds in micro propagation were investigated in various hormone treatments.
2. EXPERIMENTAL METHOD 2.1 Explants Material and Location
Explants material were collected from two years old gall rust tolerant of F. moluccana branches in one meter long. Gall rust tolerant of F. moluccana branches were collected from resistance
trial seed orchard at East Java. This study were observed in Center of Forest Biotechnology and Tree Improvement nursery at Purwobinangun, Pakem, Sleman, Yogyakarta
77.66708‘ SL and 110.42011‘ EL, 287 m asl., 26.4
C daily average temperature, 80 daily humidity, 30- 10.000 lux daily light.
2.2 Design Experiment This research was design as 2 groups of cases, branches rejuvenation in nursery and axillary
buds initiation in micro propagation. Rejuvenation as vegetative explants sources for micro propagation treatment conducted in a plastic cover nursery with crumple up system from one
meter long of two years old F. moluccana for 1 month incubation. The media of rejuvenation was top soil and sand 1:1. Rejuvenations comprise 10 families with 3 branches as replication.
Sprouting shoots from rejuvenation in each family were used for micro propagation initiation with 10 explants as replication. BAP benzyl amino purine 0.5 mgl, 1 mgl, and 1.5 mgl
used as shoots treatment and IBA indol butiric acid 1 mgl, 3 mgl and 5 mgl used as roots hormones treatment in Murashige-Skoog 1962 media. The shoot number and shoot height
per nodule were used as rejuvenation parameter, even though shoot number, shoot length and root length per explants were used as parameter in micro propagation. The research activities
need 3 months for nursery observation and 12 months for micro propagation observation.
Bogor, 21-22 October 2015
775 The group means data were analyses using post hoc tests in the Analysis of Variance
ANOVA, if significant differences exist between the treatments groups will be analysis using Duncan’s multiple range tests.
3. RESULT AND DISCUSSION 3.1 Rejuvenation of
F. moluccana
Rejuvenation of mature parts is intended to get the juvenile phase to facilitate the difficulty in the micro propagation of F. moluccana, poor reactivity of explants taken from adult trees. The
higher shoot number per nodule and height of shoots, the higher juvenile axillary bud material can collect. All branches in this research can rejuvenate the shoots in multiple number more
than 6 shoots per nodule and medium number 2-6 shoots per nodule after 1 month incubation in nursery and rooting some branch with rhyzobium nodulation after 1 year
incubation in nursery Figure 1.
a b
c d
Figure 1: Rejuvenation as vegetative explants sources conducted in a plastic cover nursery with crumple up system a, shoots from multiple nodules per branch b, shoots from single
nodule per branch c, and rooting branch with rhyzobium nodulation d. The ANOVA test appropriate for the design was carried out to detect the significance of
differences P0.05 among the treatment means Table 1. The best result for mean of shoot number 7.6667nodule ± 0.8819 was obtained in family 8 and mean of shoot height
26.5000 cmnodule ± 3.2716
was obtained in family 3. There are three groups of families in significant difference of shoot number and shoot height Table 2.
Bogor, 21-22 October 2015
776
3.2 Axillary buds initiation in F. moluccana micro propagation
Gall rust tolerant F. moluccana shoots initiation started from axillary buds after 1 week inoculation on MS solid medium supplemented with varying level of BAP. This initiation
media have good respond on sprouting ability for all F. moluccana families as explants source. In micro propagation, cytokinins include BAP are applied to promote cell division of axillary
branching. After 3 times subculture in 3 months incubation, explants shoot have 3 kind regenerated sprouting, multi sprouting more than 6 shoots, medium sprouting 2-4 shoots
and single sprouting Figure 2. The ANOVA test appropriate for the design was carried out to detect the significance of differences P0.05 among the treatment means Table 1.
Table 1: Resume of ANOVA test results for shoot number per nodule and shoots height of 10 families of F. moluccana branches rejuvenation
Source of Variation Degree of
Freedom Sum of
Squares Mean of
Square F
Sig.
shoot number Between Groups
9 76.800
8.533 4.655
0.002 Within Groups
20 36.667
1.833 Total
29 113.467
shoot height Between Groups
9 423.200
47.022 1.354
0.272 Within Groups
20 694.593
34.730 Total
29 1117.794
Significant at 95 confident interval level
a b c
Figure 2: Axillary buds regeneration in F. moluccana micro propagation with multi a, medium b and single c sprouting ability after 3 months subcultures
After 6 months subculture, the best result of mean shoot number in micro propagation was 10.00 ± 0.4472 from 0.5 mgl IBA concentration, obtain in family 8. The best result of mean
shoot length was 6.20 cm ± 0.0471 from family 2 and 6.20 cm ± 0.0422 from family 9, both were from 0.5 mgl BAP concentration Table 4. Mean of shoot number and mean of shoot
length values have not significant differences among 10 F. moluccana families, but in BAP treatments.
Bogor, 21-22 October 2015
777 Table 2: Shoot number per nodule and shoots height of 10 families of F. moluccana branches
rejuvenation for 1 month incubation in nursery
Factor Mean of shoot number
nodule Mean of shoot height
cmnodule
Family 1 2.6667 ± 0.3333
b
19.2000 ± 4.5177
b
Family 2 3.3333 ± 0.6666
b
14.3000 ± 3.6116
a
Family 3 3.0000 ± 1.0000
c
26.5000 ± 3.2716
c
Family 4 3.6667 ± 1.4530
b
25.6667 ± 4.2463
b
Family 5 3.0000 ± 0.0000
b
21.1000 ± 5.4745
b
Family 6 1.3333 ± 0.3333
a
24.4000 ± 0.9504
b
Family 7 4.3333 ± 08819
c
19.6667 ± 0.6333
b
Family 8 7.6667 ± 0.8819
d
17.5667 ± 3.7391
b
Family 9 4.0000 ± 0.0001
c
19.3333 ± 0.5487
b
Family 10 2.3333 ± 0.8819
b
17.0333 ± 2.8322
b
Values average ± standard error with different letters are statistically significant according to Duncans multiple comparison tests
Table 3: Resume of ANOVA test results of effect of various BAP in MS medium on shoot number and shoot length of F. moluccana axillary buds initiation
Source of Variation Degree of
Freedom Sum of
Squares Mean of
Square F
Sig.
shoot number Between Groups
9 76.800
8.533 4.655
0.002 Within Groups
20 36.667
1.833 Total
29 113.467
shoot length Between Groups
9 423.200
47.022 1.354
0.272 Within Groups
20 694.593
34.730 Total
29 1117.794
Significant at 95 confident interval level
Table 4: Effect of various BAP in MS medium on shoot number and shoot length regeneration of F. moluccana micro propagation after 6 months subculture
Factor Mean of shoot number
nodule Mean of shoot length
cmnodule BAP concentration mgl
0.5 1
1.5 0.5
1 1.5
Family 1 6.70 ± 0.3958
a
3.30 ± 0.4485
b
1.40 ± 0.2211
c
5.60 ± 0.0650
d
5.10 ± 0.0211
e
4.82 ± 0.0442
f
Family 2 6.70 ± 0.6506
a
3.10 ± 0.3480
a
1.80 ± 0.3887
b
6.20 ± 0.0471
d
5.00 ± 0.0633
e
4.74 ± 0.0805
f
Family 3 7.00 ± 0.5773
a
3.20 ± 0.4667
b
1.50 ± 0.2236
c
6.10 ± 0.0258
d
4.80 ± 0.0422
e
4.40 ± 0.0667
f
Family 4 6.00 ± 0.4888
a
3.60 ± 0.5416
b
1.70 ± 0.2603
c
5.50 ± 0.0954
d
4.20 ± 0.0615
e
3.80 ± 0.0615
f
Family 5 6.90 ± 0.5167
a
3.00 ± 0.3652
b
1.40 ± 0.2211
c
5.82 ± 0.0359
d
4.20 ± 0.0615
e
3.80 ± 0.0615
f
Family 6 7.10 ±0. 4069
a
2.80 ± 0.4163
b
1.50 ± 0.2236
c
5.80 ± 0.0211
d
4.00 ± 0.0447
d
4.00 ± 0.0558
e
Family 7 6.70 ± 0.4485
a
3.10 ± 0.4069
b
1.30 ± 0.1528
c
6.10 ± 0.0218
d
4.00 ± 0.0447
d
4.00 ± 0.0558
e
Family 8 10.00 ± 0.4472
a
6.50 ± 0.5426
b
3.00 ± 0.4472
c
6.00 ± 0.0422
d
4.00 ± 0.0447
e
3.60 ± 0.0103
f
Family 9 6.70 ± 0.6155
a
3.80 ± 0.5121
b
1.10 ± 0.1000
c
6.20 ± 0.0422
d
5.00 ± 0.0333
e
3.90 ± 0.0258
f
Family 10 6.00 ± 0.6831
a
3.20 ± 0.5121
b
1.50 ± 0.3073
c
5.10 ± 0.0258
d
5.00 ± 0.0333
e
3.80 ± 0.0615
e
Values average ± standard error with different letters are statistically significant according to Duncans multiple comparison tests.
Bogor, 21-22 October 2015
778 Figure 3: Root length of 10 families of F. moluccana with IBA treatments in MS media
after 12 months subcultures After 12 months subculture, the best result of mean root length was obtained from 3 mgl
IBA concentration treatment Figure 3. The concentration response curves of plant hormones are generally bell-shaped. At a too low concentration there is no effect, and at a too
high concentration the added hormone is inhibitory. The optimum effect only occurs at intermediate concentrations. The formation of roots from pith fragments corresponds with
the effect of auxin on rooting of cuttings, and the reduction of shoot formation with the inhibition of the outgrowth of axillary buds by auxin George Debergh, 2008. The
epidermis of plants is relatively impermeable to auxin and most uptakes by explants occur via the cut surface Taiz, 2006. Roots are formed from founder cells close to the cut ends so
auxin may reach these cells by diffusion.
4. CONCLUSION
In the present study, rejuvenation of branch as explants source in micro propagation were investigated for the establishment of vegetative micro propagation protocol efficiently in gall
rust tolerant Falcataria moluccana. Rejuvenation with crumple up system provides useful technique for F. moluccana to get the juvenile phase to facilitate the difficulty in poor reactivity
of explants taken from adult trees. The best results of rejuvenation and micro propagation with multi shoot sprouting ability were obtained in family 8. After 6 months micro
propagation subculture, 0.5 mgl BAP in MS media give the best result for axillary buds initiation and after 12 months subculture, 3 mgl IBA in MS media give the best result for
roots formation.
ACKNOWLEDGMENT
The author would like to thank Toni Herawan, Fithry Ardhani, Titis Budi Ekowati as researcher team for helpful discussions. We are grateful to Suprihati, Endin Izudin, Rudi
Hartono and Waluyo for technical assistants.
M ean
of m
ic ro p
rop a
g at
ion
root l
en g
th
F. moluccana families 1 mgl IBA
3 mgl IBA 5 mgl IBA
Bogor, 21-22 October 2015
779
REFERENCES
Ballester, A., San-Jose M. C., Vidal N., Fernandez-Lorenzo J. L., Vieitez A. M. 1999. Anatomical and Biochemical Events during the Rooting In Vitro of Microcuttings from
Juvenile and Mature Phases of Chesnut. Annals of Botany 83:619-629. Basheer, R., Salimia. 2007. Juvenility, Maturity and Rejuvenation in Woody Plants. Hebron
University Research Journal. Vol 3, No. 1. pp 17-43. Bon, M.C., Bonal D., Goh, D., Monteuuis, O. 1998. Influence of Different Macronutrient
Solutions and Growth Regulators on Micropropagation of Juvenile Acacia mangium and Paraserianthes falcataria explants. Plant Cell, Tissue and Organ Culture 53: 171-177.
Chujo, M., Eizawa, J., Yokota, S., Ishiguri, F., Lizuka, K., Priadi, D., Sumiasri, N., Yoshizawa, N. 2010. Micropropagation and Protoplast Culture in Paraserianthes
falcataria. Wood Research Journal Vol.1, No. 2. George, E.F., Debergh P.C. 2008. Micropropagation: Uses and Methods In Plant
Propagation by Tissue Culture 3rd Edition. E.F. George, M.A. Hall G-Jan De Klerk eds.. Springer Pub. Netherland.
George, E.F., de Klerk, G.J. 2008. The Components of Plant Tissue Culture Media I: Macro-and Micro-Nutrients In Plant Propagation by Tissue Culture 3
rd
Edition. E.F. George, M.A. Hall and Geert-Jan de Klerk eds. Volume 1. The Backforund. Springer
Pub., Netherland. Libby, W.J., Brown, A.G., Fulding, J.M. 1972. Effects of Hedging Radiata Pine on
Production, Rooting and Early Growth of Cutting. New Zealand J. For. Science. 2:263- 283.
Machakova, I., Zazimalova, E., George, E.F. 2008. Plant Growth Regulators I: Introduction; Auxins, their Analogues and Inhibitors In Plant Propagation by Tissue
Culture 3
rd
Edition. E.F. George, M.A. Hall and Geert-Jan de Klerk eds. Volume 1. The Backforund. Springer Pub., Netherland.
Murashige, T., Skoog, F. 1962. A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Culture. Physiol. Plant. 15:473-497.
Nehra, N.S., Becwar, M.R., Rottmann, W.H., Pearson, L., Chowdhury, K., Chang, S., Wilde, H. D., Kodrzycki, R.J. 2010. Wood Research Journal Vol.1, No. 2.
Sukarutiningsih, Saito Y., Ide Y. 2002. In Vitro Plantlet Regeneration of Paraserianthes falcataria L. Nielsen. Bulletin of The Tokyo University Forests 107: 21-28.
Sumiasri, N., Priadi, D., Yokota, S., Yoshizawa, N. 2006. Tissue Culture of Fast Growing Tropical Trees in Indonesia: Mangium Acacia Mangium Wild and Sengon Paraserianthes
falcataria L Nielsen. In: Imamura, Y.; T. Umezawa; T. Hata Eds. Sustainable Development and Utilization of Tropical Forest Resources. Research Institute for
Sustainable Humanosphere, Kyoto University, Kyoto, P. 123-130.
Taiz, L., Zeiger, E. 2006. Plant Physiology. Sinauer Associates Inc, Sunderland. Wróblewska, K. 2013. Benzyladenine Effect on Rooting and Axillary Shoot Outgrowth of
Gaura Lindheimeri Engelm. A. Gray Cuttings. Wrocław University of Environmental
and Life Sciences. Acta Sci. Pol., Hortorum Cultus 123 2013, 127-136.
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780
POSTER F13 - Alternative Propagation of Petung Bamboo
Dendrocalamus asper
Schultes f. Backer ex Heyne without Taking Apart of Clump
Merryana Kiding Allo
1
Forestry Research Institute of Makassar, Jl. Perintis Kemerdekaan km. 16 Makassar, 90243 Corresponding Email: merrymksryahoo.co.id; merrymakassargmail.com.
ABSTRACT
Petung bamboo is the largest diameter of bamboo species found in Indonesia. The strength and durability of the fiber of Petung bamboo have been tested so many use full for bridge
construction, building houses, butterfly fish in the sea, furniture and many other uses. The necessary cultivation techniques of Petung bamboo is too important for availability of raw
materials of bamboo. Assuming some people are very difficult and requires a high cost for multiply bamboo, because to be done by cuttings rhizome so it must unload clumps that can
be deadly clumps. The yield of research found a way to multiply the twig of branch, by soaking for 2 weeks - 1 month with the addition of liquid fertilizer in the shade up to 70
yield up to 57.77 growth.
Keywords: Petung bamboo, reproduction, twig. 1. INTRODUCTION
Petung bamboo in Indonesia is well known about the strength of the fiber and it’s durable.
Natural stands of bambu cannot be fulfilled as well as bamboo forests of the people so as to support the industry required the development of bamboo plantations managed
professionally. Strength fiber of bamboo rods approach the strength of the wood so as wood substitute material substitution, in addition to the bamboo fiber patterns petung pertained
beautiful species of bamboo veneer for the material. Have a higher rate than the growth rate of trees. Another utilization of the bamboo shoots petung is as a producer of well-loved flavor
with a smooth texture and sweet enough remains favoured by consumer food bamboo shoots in Asian countries.
Petung bamboo Dendrocalamus asper Schultes f. Backer ex Heyne is the largest diameter of bamboo species diameter 25 centimetres, the segments of rod is 31 and getting longer up
to 26 meters. The economic and ecological value of petung bamboo plants highest among of the other bamboo species. The root system have symposia, very strong, growing tightly,
spread in all directions and to produce solid clumps as windbreaks plant. Another utilization of petung bamboo as a producer bamboo shoots were very popular in Asian countries
because of the taste, texture quite smooth and contains high fiber. The ecological function, such a stimulation of above ground andor below ground biomass growth could potential
produce more renewable energy and more C benefits to project proponents in CDM or REDD, the results appear lacking, and variables beyond biomass productivity determine
ecosystem or biome C stocks and flows Důking et al. 2011. In the generally, bamboo in Indonesia have develop themselves with vegetative through
rhizomes as a place to grow young shoots candidates rod, so that the development cannot be expected to grow in large numbers. Rarely found the bamboo farmers or land owners will try
Bogor, 21-22 October 2015
781 to plant itself, just expected growth naturally. Lack of public awareness in the cultivation of
bamboo raises concerns about the extinction of these kinds of bamboo. Because the older of the bamboo grove petung diminishing productivity may even die before flowering and fruiting
due to disease, as happened in 1998 in Thailand. One of the problems of bamboo cultivation in Indonesia is preparation of seed, the generally
made at the expense of productive bamboo grove and the result of the multiplication is not as expected. While attempts to wait bamboos flower and produce fruit and seeds can only occur
once during the cycle of a clump. Similarly, by developing from seeds or shoots through tissue culture requires a high cost to a long time Sutyono, 1996.
Conceivably, if you needs the bamboo seeds, you must ruin the clump will be sacrificed and how much manpower is needed for the job?
In order for the preservation of bamboo petung can continue, it takes the bamboo cultivation techniques are easy and cheap to make it affordable for people in rural practice in order to
meet the needs of both local and national.
2. EXPERIMENTAL METHOD 2.1 Location and time schedule
This study was carried out in the nursery Bambusetum Forest Area Special Purpose KHDTK Mengkendek, Tana Toraja, South Sulawesi Province. The research was conducted during the
10 months from September 2014 - June 2015.
2.2 Equipment and Materials Materials used are branch cuttings of petung bamboo in Mengkendek KHDTK crops in 2001,
manure and soil media. The equipment used is whittle, saws, shovels, pesese, poly bags, gloves, footwear field, label, counter for counting the number of shoots appearing, growth
measurement tools such as rollers meter and a ruler to measure the height of the young shoots and stems of bamboo.
2.3 Implementation Research Preparation of the planting medium, made after 4 weeks the plant material is placed in damp
areas close to water sources. Making the planting medium by mixing 3 parts soil to 1 part manure that has been cooked through drying to prevent mold growth in high humidity. Stir
mixture of soil and fertilizer and enter into a polybag until filled ¾ part, after the completion polybag filled all let stand for a week in order to land in it solidified and meetings by means of
flushing water every 2 days. Put polybag in the shade under humid conditions 68-78. Selection of plant material taken from a branch stem attached to each segment to 5-14, 15-
20 and 21-28 and diameter measure 1.5 to 2.2 cm. Branch cuttings are made by removing the branches from stem segments using a machete carefully by means of cutting on the stump
until detached, pruning entire branches from cuttings taken leafy branches up to 5 cm of branch weevil round.
Storage, put the plant material in a place that is always moist but not to be submerged, cover with bamboo leaves to maintain moisture. Within 4-6 weeks shoots will be visible candidates
will appear prominently on the stump section yellowish-white twig.
Bogor, 21-22 October 2015
782 Observations of growth, started in candidate shoots appear in the place of storage, subsequent
developments noted in particular a period of up to sprout. Weaning the roots grow into a polybag after soaking for 30 minutes with a role in root growth
hormone solution that can root growth expansion. 2.4 Method of collecting data
Observations and direct measurements of properties such as the number of candidates is growing shoots and shoots that appear at the root nodules. These observations were made at
the time of the cuttings placed in a high humidity while waiting for roots to grow. While the growth of branches, young shoots of bamboo and a high stem density observed after the
cuttings are planted in polybasic. Other primary data is from the nursery include measurements of air temperature, humidity and radiation. Secondary data is needed is a
common condition of study sites, among others, geography, topography, climatic conditions, soil conditions, management of bamboo stands history and socioeconomic conditions settler
communities around KHDTK Mengkendek. Data analysis using MS excel, according to data the percentage growth, the number of shoots
and shoot height and number of shoots in time and presentation of test results done in the form of descriptive discussion.
3. RESULT AND DISCUSSION 3.1 Growth
