ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 In Vitro Propagation of Albizia Lebbeck Through Axillary Bud Culture K.R. Al-Joboury Iraq Natural History Research Center & Museum,Unive rsity of Baghdad Received in : 20 April 2011 Accepte d in : 16 November 2011 Abstract The p resent st udy describes a p rotocol for rapid in vitro microprop agation of Albizia lebbeck during the p eriod of October 2007 to October 2009 through nodal segments containing axillary buds. The buds induced to p roduce a large number of multiple shoot s by culturing on M S medium sup p lemented with different concentrations of BA (benzy ladenine) and NAA (α-nap hthalene acetic acid). The maximum number of shoot s p er exp lants was obtained on M S medium sup p lemented with 1.0 mg/L BA and 0.1 mg/L NAA was (4.8) after 4 weeks of culture. Excised shoot s were rooted on half strength M S medium fortified with 0.5 mg/L either IBA (indolbuty ric acid) or NAA alone. The comp lete plantlets t hus obt ained were successfully transferred to soil. Key words: Albizia lebbeck, tissue culture, in vitro, p lant regeneration Indroduction The genus Albizia (also Albizz ia) Albizia lebbeck (L.) Benth. commonly known as Siris is a mimosoid tree legume widely distributed in the world. belongs t o subfamily M imoseae of family Leguminosae (Fabaceae ) / Pea Family and is highly valued multip urp ose tree legume. [1,2] . The genus Albizia consists of app roximately 150 sp ecies distributed in Asia , Africa , Australia , and trop ical and subt rop ical America Several Albizia sp ecies are p lanted as ornamentals or as a source of tannin extracts [3] Albizia lebbeck is a deciduous woody trees and shrubs and forms sy mbiotic relationship with Rhizobium and fix atmosp heric nitrogen used for its growt h and also for the enrichment of the rhizosp here [4] One of the most imp ortant p roblems in the woody legumes in the establishment of p rop agules of high quality of regeneration as Albizia lebbeck has long seed dormancy. Under tissue culture app roach the p lant sp p belonging to the family leguminoseae are in general considered to be recalcitrant to in vito regeneration. under these efforts Albizia lebbeck was subjected to tissue culture technique for developing the avenues for mass p roduction and development of elite genoty p e of Albizia lebbeck from diggerent exp lants. [5] It is thus necessary to device a rapid and efficient microprop agation p rotocol for obtaining true‐to‐ty p e regenerants without detriment to the survival of mother/donor p lant and saving its p op ulations from gett ing rarer in nature. aiming to st udy the microprop agation of Albizia lebbeck to solve some of the prop agation p roblems in this p lant. ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Material and Methods Shoot apices having nodal segments with axillary buds were collected from natural trees growing in Al-Rashdia area. The buds were disinfected with 0.5% sodium hy p ochlorite solution with 3-5 drop s of T ween- 20 for 20 minutes and washed thoroughly with tap water. The buds were also treated with 0.05% mecuric chloride solution for 5 minutes then washed with st erile distilled water and p retreated with citric acid (150 mg/l), ascorbic acid ( 100 mg/l ). The st erilized nodal exp lants were transferred to M urashige and Skoog (1962) (M S) medium sup p lemented with growt h regulators at various concentrations and combinations of BA with NAA for shoot induction. Excised shoot s from these cultures were rooted on ½ M S sup p lemented with IBA (or NAA at different concentrations for root induction p H was adjust ed to 5.6 using 0.1 N NaOH or 0.1 N HCl before autoclaving ( 121°C under 1.05 kg/cm2, 20 min ). The cultures were maintained in a culture room at 25±2°C and were exp osed to continuous fluorescent light for 16 h p er day and successfully transferred to soil. Significant differences among mean values were using One-way ANOVA followed by Duncan's multiple-range test was conducted to evaluate differences among the treatments. Re sults and Discussion M icrop rop agation is the true-to-t y p e p rop agation of selected cultivar using in vitro technique. The segments of Albizia lebbeck exhibited browning of the exp lant and medium due to leaching of p henolics. This p henomenon results from phy siological changes within the cultured tissues that lead to gradual browning and eventual death of t issues. [6] Tissue browning is a p roblem frequently observed during in vitro establishment of exp lants from woody p lants [7]. The p roblem of phenolic browning was minimized to a great extent by leaching of p henolic comp ounds due to agitation in antioxidants solution and by p rop er dry ing of exp lants p rior to inoculation [8] Significantly reduced leachin by sup p lementing the medium with citric acid (150 mg/l), ascorbic acid ( 100 mg/l ) This was in agreement with [9]. The variance analysis of the axillary bud cultures showed that the effects of the treatments with different combination of BA and NAA were significant on number of shoot . The nodal segments p roved to be excellent exp lants for multiple shoot formation and the first resp onse of axillary buds within two weeks. New shoot development from axillary bud was observed within three weeks of culture and more shoot s were found to develop during subcultures. The best resp onse was found under 1.0 BA+0.1 NAA mg /L combination which was found most effective (Fig. 1). The combined effects of BA+NAA on shoot induction were reported earlier in different p lants. Our results agreed with those obtained by [10,11,12]. On the other hand the adding of NAA in different concentrations developed the multiple shoot s in this st udy but the best concentration was 0.6 mg/l. Root formation was induced when elongated shoot s (1-2 cm) were transferred to ½ M S medium fortified with 0.5 mg/L IBA or NAA . The result showed that t he IBA is considered as t he most effective auxin in root induction. Elongated shoot s derived from the exp lants were separated and cultured on half st rength M S sup p lemented with IBA (0.5 mg/l) for induction of rooting. (Fig. 2) The op timum concentration was 0.5 mg/l of IBA and it resulted in 85% of root initiation within 1- 2 weeks of culture[13,14] The comp lete p lantlets were transferred to small plast ic p ots, and bags containing a mixture of soil and compost then transferred to t he field. (Fig. 3) ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 Re ferences 1-Saha, A. and Ahmed, M . (2009). The analgesic and anti-inflammatory activities of the extract of Albizia lebbecl in animal model . Pak. J. Pharm. Sci., 22(1): 74-77 2- Ghary al. P. and M aheshwari, S. (1983) In vitro differentiation of p lantlets from tissue cultures of Albizia lebbeck L. Plant Cell Tissue Organ Culture 2:49-53 3-Seyy ednejad, S. ; Niknejad, M . and Yusefi, M . (2009). Study of air p ollution effects on some p hy siology and morphology factors of Albzia lebbeck in high temp erature condition in Khuz estan. J. Plant Sciences, 4: 122-126 4-Qadri, R. and M ahmood, A. (2005) . Ultra-Structural st udies on root nodules of Albezia lebbeck (L.) Benth. J. Bot ., 37(4): 815-82 5-M amun, A. ; M atin, M . ; Bari, M . ; Siddique, N. ; Sultana, R. ; Rahman, M . and M usa, A. (2004). M igrop agation of woody legume (Albizia lebbeck ) through tissue culture. Pakist an J. Biol. Sci., 7(7): 1099-1103 6-Alkhateeb, A. and Ali-Dinar, M . (2002). date p alm in Kingdom of Saudi Arabia: cultivation, p roduction and p rocessing. translation, authorship and p ublishing center, King Faisal University , Kingdom of Saudi Arabia: 188. 7-Block, R. and Lankes, C. (1996). M easures to p revent tissue browning of exp lants of the app le rootst ock M 9 during in vitro establishment. Gartenbauwissenschaft, 61:11-17. 8-M eghwal, P. ; Sharma, H. and Singh, S. (2001) Effect of surface st erilizing agents on in vitro culture establishment of guava (Psidium guajava L.). Progressive Hort iculture, 33:101- 103. 9-Badawy , A. ; Habib, A. ; El-Ban, A. and Yosry , G. (2005). Prop agation of Dracaena fragrans p lants by tissue culture technique. Arab J. Biotech., 8 (2) : 329-342. 10-Lal, N. and Ahuja, P. (2000). Adventitious shoot bud formation from cultured leaf exp lants of Rheum emodi Wall. Plant T issue Cult., 10: 17–24 11-M unshi, M . ; Hakimi, L. ; . Islam, A. and Ahmed, G. (2004). In vitro clonal p rop agation of banyan (Ficus benghalensis L.) through axillary bud culture . Int. J. Agri. Biol., 6 ( 2 ) 12-Al-Sulaiman, M . and Barakat, M . (2010). In vitro shoot multiplication of Ziziphus spina- christi by shoot tip culture. J. Bio., 9(6): 850-857 13-Dhabhai, K. ; Sharma, M . and Batra, A. (2010). In vitro clonal p rop agation of Acacia nilotica (L.) - A nitrogen fixingtree. Researcher, 2010, 2(3) 14-Tomar, U. and Gup ta, S. (1988). In vitro p lant regeneration of leguminous trees (Albizia sp p .). Plant Cell Reports, 7: 385–388 ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 0 20 40 60 80 100 0.5+0.05 1.0+0.10 1.5+0.5 concentrations of BA+NAA % r e s p o n s e 0 2 4 6 n o m b e r o f s h o o ts p e r e x p la n ts Fig (1): Effect of BA and NAA concentrations on shoot inducation of Albizia lebbeck at 4 weeks . 0 2 4 6 8 10 N o . ro o ts p e r e x p la n ts 0.1 0.5 1 concentration of IBA and NAA alone IBA NAA Fig( 2): Effects of different concentrations of IBA and NAA on rooting of excised shoot in Albizia lebbeck ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 A- B- Fig(3): In vitro p rop agation of Albizia lebbeck A-root ing of excised shoot in ½ M S medium sup p lemented with 0.5 IBA (mg/L), B- establishment of p lants in p last ic p ot ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012 اإلكثار خارج الجسم الحي لـ Albizia lebbeckبوساطة البراعم االبطیة خنساء رشید الجبوري جامعة بغداد/ مركز بحوث ومتحف التاریخ الطبیعي 2011 تشرین الثاني 16:قبل البحث في ،2011 نیسان20: استلم البحث في الخالصة المــدة مـــن Albizia lebbeck بیـــنت هــذه الدراســة طریقـــة اإلكثــار الخــضري الــسریع خـــارج الجــسـم الحـــي لـــ باستعمال قطع عقدیة حاویـة بـراعم ابطیـة التـي اسـتحثت إلنتـاج اكبــر عـدد مـن الـسیقـان 2009 الى أكتوبـر – 2007أكتوبر وتـم الحـصول علـى اكبـر عـدد مـن . بـصورة منفـردةNAA و BAٍمن خالل زراعتها في وسط حاو على تراكیـز مختلفــة مـن ، ثم جذرت هذه 4.8ن عددها ، إذ كاNAAلتر / ملغم0.1و BA لتر / ملغم 1.0النمـوات في وسط حـاوي على تركیز Mٍالنمـوات فــي وســط حــاوعلى نــصـف القـوة مــن الوســـط S مــضـافا إلیـــه IBA لتـــر ثـــم نقلــت النمـــوات / ملغــم 0.5 بتركیـز .الناتجـة بصورة ناجحة إلى التربة ، زراعة االنسجة ، خارج الجسم الحي ، إكثار النباتاتAlbizia lebbeck: الكلمات المفتاحیة ة مجلة إبن الھیثم للعلوم الصرفة و التطبیقی 2012 السنة 25 المجلد 1 العدد Ibn A l-Hai tham Journal f or Pure and Applied Science No. 1 Vol. 25 Year 2012