茵芋花粉培养效应的优化
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摘要
为研究茵芋花粉的培养效应,采用蔗糖、硼酸、氯化钙三因素三水平培养基离体培养法,I2—KI、TTC、红墨水等染色法,以及生物荧光显微镜对花粉活力进行观察与检测,筛选有活力的花粉的最适培养基及活力测定方法.结果表明,新鲜茵芋花粉粒呈黄色,花粉离体后逐渐转变为黄褐色.I2—KI、TTC对花粉染色无效;红墨水染色法使有活力的花粉在蓝色滤光片下呈绿色或原色,能够快速有效地检测花粉活力.花粉于25℃的恒温下离体培养4 h开始萌发,至24 h萌发稳定.蔗糖、硼酸、氯化钙三因素三水平培养基离体培养对花粉萌发的互作效应极显著,最优培养基为:15 g·L~(-1)蔗糖+0.05 g·L~(-1)硼酸+0.02 g·L~(-1)氯化钙,萌发率为60.0%;次优培养基为:15 g·L~(-1)蔗糖+0.03 g·L~(-1)硼酸+0.02 g·L~(-1)氯化钙,萌发率为58.0%;第三培养基为:15 g·L~(-1)蔗糖+0.01 g·L~(-1)硼酸+0.01 g·L~(-1)氯化钙,萌发率为50.4%.影响茵芋花粉萌发的主因素为氯化钙,次因素为蔗糖,辅助因素为硼酸.
In order to screen out the optimal medium for in vitro pollen germination of Skimmia reevesiana and reliable pollen vitality evaluation method,an orthogonal design with 3 factors in 3 levels,including sucrose,boric acid and calcium chloride,were implemented. Pollen vitality was evaluated by methods of I2—KI,TTC,red staining,and bioluminescent display. The results showed that fresh S.reevesiana pollen was yellow and turned yellowish brown in vitro. I2—KI and TTC methods were inapplicable to vitality test.However,by red ink staining,vital S.reevesiana pollen was green or in the original color under blue optical filter,indicating that it is an effective method for pollen vitality test. Pollen began to germinate 4 h after in vitro culture at 25 ℃ and reached stable status within 24 h. The interactive effect of 3 chemicals on in vitro pollen germination was significant. The optimal medium formula was 15 g·L~(-1) sucrose,0.05 g·L~(-1) boric acid,and 0.02 g·L~(-1) calcium chloride,reaching a 60.0% germination rate. Among all factors that affecting germination rate,calcium chloride was the predominate factor,which was followed by sucrose and boric acid.
In order to screen out the optimal medium for in vitro pollen germination of Skimmia reevesiana and reliable pollen vitality evaluation method,an orthogonal design with 3 factors in 3 levels,including sucrose,boric acid and calcium chloride,were implemented. Pollen vitality was evaluated by methods of I2—KI,TTC,red staining,and bioluminescent display. The results showed that fresh S.reevesiana pollen was yellow and turned yellowish brown in vitro. I2—KI and TTC methods were inapplicable to vitality test.However,by red ink staining,vital S.reevesiana pollen was green or in the original color under blue optical filter,indicating that it is an effective method for pollen vitality test. Pollen began to germinate 4 h after in vitro culture at 25 ℃ and reached stable status within 24 h. The interactive effect of 3 chemicals on in vitro pollen germination was significant. The optimal medium formula was 15 g·L~(-1) sucrose,0.05 g·L~(-1) boric acid,and 0.02 g·L~(-1) calcium chloride,reaching a 60.0% germination rate. Among all factors that affecting germination rate,calcium chloride was the predominate factor,which was followed by sucrose and boric acid.
引文
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[2]郑天汉,吴远彬,阮淑明,等.茵芋生物学特征观测[J].武夷学院学报,2016,35(12):31-33.
[3]郑天汉,李勇,吴远彬,等.茵芋驯化栽培与植物生长调节剂促进效应研究[J].天津农业科学,2016,22(8):125-129.
[4]郑天汉,张志才,李勇,等.基质与激素对茵芋扦插萌芽效应研究[J].山东林业科技,2016,46(5):23-28.
[5]范宝磊,张耀兮,吴仲,等.不同品系油菜花粉生活力测定方法比较[J].安徽农业科学,2012,40(5):2 596-2 597.
[6]赵鸿杰,乔龙巴图,殷爱华,等.3种山茶属植物花粉活力测定方法的比较[J].中南林业科技大学学报,2010,30(3):105-107.
[7]袁德义,谭晓风,邹锋,等.油茶花粉生活力检测方法比较[J].西南林学院学报,2009,29(4):10-12.
[8]胡适宜.植物胚胎实验方法(一)花粉生活力的测定[J].植物学通报,1993,10(2):60-62.
[9]任芳.连翘组织培养、花粉活性和雌雄配子体发育研究[D].太原:山西大学,2006.
[10]程伟.西南桦花粉物候、生殖构件和花粉特性研究[D].南宁:广西大学,2007.
[11]张建浩,吴永美.红墨水(酸性大红G)染色法测定大豆生活力[J].农业技术与装备,2009(6):62.
[12]胡春,刘左军,李富香,等.钝裂银莲花花粉活力测定方法的研究[J].植物研究,2013,33(5):582-586.
[13]李畅,苏家乐,刘晓青,等.一品红27个品种花粉量、花粉活力及3种测定方法的比较[J].江西农业大学学报,2012,34(6):1 130-1 135.
[14]赵统利,周翔,朱朋波,等.百合花粉生活力测定方法的比较研究[J].江苏农业科学,2006(5):88,144.