歐洲丁香品種‘Downfield’花序和花序軸愈傷組織誘導(dǎo)和懸浮培養(yǎng)1)

于秋瑩 郭苗苗 許岢昕 劉建斌 王聰 鄭健 張克中 張炎

(北京農(nóng)學(xué)院，北京，102206)

The inflorescences and inflorescence axial of Syringa vulgaris as explants were used to study the effects of explant types, basic culture medium types, plant hormone types and concentration to the callus induction rate of S. vulgaris, and design orthogonal experiment was used to explore the effects of erlenmeyerflask volume, initial culture medium volume, retention of old medium, inoculum quality and the content of antioxidants to the callus proliferaton result of suspension culture. The optimal callus induction of inflorescence axis medium was modified as Blaydes+TDZ 0.10 mg/L+2,4-D 1 mg/L, the optimal inflorescence induction medium was MS+TDZ 0.10 mg/L+2,4-D 1 mg/L, the induction rate was up to 100%. The concentration of 2,4-D had significant effect the callus rate of inflorescence and inflorescence axis (p<0.05). The combination of TDZ and 2,4-D significantly affected the inflorescence axial callus rate (p<0.05). The optimal combination of suspension culture was using the volume of 250 mL erlenmeyer flask, and supplemented with 150 mL of initial culture mediumaccompanied by 4.5 g of calli, andafter 15 d, retained of 1/4 old medium for next stage, initial inoculumed 7 g, PVP concentration 100 mg/L, Vc concentration 75 mg/L. This is the first time to report callus induction and start cell suspension culture of inflorescence and inflorescence axis of S. vulgariswith successful results, which would be used for factory breeding and genetic transformation in the future.

歐洲丁香(Syringavulgaris)是木樨科(Oleaceae)丁香屬(Syringaspp.)知名的觀花灌木或小喬木[1]，具有觀賞、經(jīng)濟(jì)[2]、生態(tài)[3]等多種價(jià)值，是目前應(yīng)用最為廣泛的丁香屬樹(shù)種。隨著歐洲丁香需求量不斷增大，如何提高其繁殖效率受到廣泛關(guān)注。目前對(duì)于繁殖木本優(yōu)良品種，只能采用無(wú)性繁殖的方法，其中，組織培養(yǎng)技術(shù)較其他傳統(tǒng)的無(wú)性繁殖技術(shù)具有繁殖系數(shù)大、繁殖速度快、不受季節(jié)影響等優(yōu)點(diǎn)[1]，并且被逐漸應(yīng)用于丁香。國(guó)內(nèi)外已經(jīng)對(duì)紫丁香(SyringaoblataLindl.)[4]、暴馬丁香(Syringareticulatavaramurensis)[5]、歐洲丁香[6]等多種丁香的組培方法進(jìn)行研究，主要開(kāi)展了離體胚培養(yǎng)[7]、腋芽增殖培養(yǎng)[6]、愈傷組織誘導(dǎo)[8]、體細(xì)胞胚胎發(fā)生[9]等研究。愈傷組織的誘導(dǎo)是離體再生的關(guān)鍵一環(huán)，影響愈傷組織誘導(dǎo)的因素有外植體類(lèi)型[10]、培養(yǎng)基種類(lèi)[11]、激素種類(lèi)及濃度[12]等，其中外植體類(lèi)型的影響很大，目前在丁香屬植物愈傷組織誘導(dǎo)中，多使用葉片、莖段、上胚軸等營(yíng)養(yǎng)器官作為外植體[12-14]，尚未見(jiàn)采用花序及花序軸等生殖器官進(jìn)行誘導(dǎo)的報(bào)道。有研究表明，花序及花序軸作為生殖器官較營(yíng)養(yǎng)器官更容易產(chǎn)生體細(xì)胞愈傷組織和體細(xì)胞胚胎[15-16]。愈傷組織具有多種用途，可以作為建立離體再生、遺傳轉(zhuǎn)化體系和提取次生代謝產(chǎn)物的材料[17-19]。與傳統(tǒng)的固體培養(yǎng)基培養(yǎng)相比，液體懸浮震蕩培養(yǎng)是一種高效的愈傷組織增殖培養(yǎng)方法[20]。由于愈傷組織在液體中與營(yíng)養(yǎng)物質(zhì)的接觸面更大，且震蕩使其分離成單細(xì)胞，更易分離和生長(zhǎng)，從而提高愈傷組織增殖速率[21]?！?br>