长寿花（Kalanchoe blossfeldiana V. Poelln.）试管微嫁接技术的系统研究
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摘要
长寿花(Kalancho eblossfeldiana V.Poelln.)是景天科(Crassulaceae.)伽蓝菜属(KalanchoeAdans.)多年生肉质的观叶和观花植物,在盆栽花卉中占有重要的地位,是国际花卉市场中发展最快的盆花之一。国内对长寿花的研究到目前为止主要是关于长寿花栽培、繁殖技术和组织培养方面的内容,国外多集中在生理因素和内含物方面的研究。而国内外对于品种选育的研究较少,且现有品种已经无法满足市场品种更新的要求。利用试管微嫁接技术是进行品种改良的有效途径之一,但目前尚未见对于长寿花的试管微嫁接技术及影响其成活率的因素方面的系统研究报道。
本试验从六个长寿花品种中(C1～C6)选择了叶片形态与花色差异较大的两个长寿花品种C3(紫缤)和C6(未知)作为试验材料,以C3为接穗,C6为砧木。利用已经建立的尢菌体系,从嫁接方法、培养条件、砧木接穗的处理等方面对微嫁接技术体系进行了系统研究,主要研究结果如下:
利用原有的长寿花无菌快繁体系能达到快速高效的繁殖目的,但不适宜作为砧木使用。将品种C6的再生植株在MS+GA0.5mg/L+BA1.0mg/L的培养基中培养30d,得到节间伸长均匀,且平均茎粗为0.179cm,平均株高7.6cm的不定芽,适宜作为微嫁接的砧木。
嫁接方法采用劈接法和顶接法,结果表明:顶接法的嫁接成活率为73.3%,而劈接法的成活率仅为33.3%,因此,认为长寿花试管微嫁接的适宜方法为顶接法。
激素对长寿花试管微嫁接苗成活率的影响方面,研究了在MS培养基中添加不同浓度的BA、IAA、NAA,结果表明:加入IAA的培养基中嫁接苗的成活率为0,说明IAA不利于嫁接苗的成活。在培养基MS+BA1.0mg/L+NAA0.1mg/L中,嫁接的成活率为100%,平均株高1.1cm,生长速率最快为0.009cm/d,嫁接苗的接穗和砧木生长状态良好,基本上没有黄叶和落叶的现象,生长均匀,生长速率较快,茎段的粗度有所增加。因此,适于长寿花嫁接苗快速生长的最佳培养基为MS+BA1.0mg/L+NAA0.1mg/L。
暗培养10d、15d、20d与对照(光培养)相比较的结果:暗培养20d的成活率为0.15d的成活率仅63.3%,而光培养和暗培养10d的成活率达到100%,且暗培养10d时,大部分接穗都有生长的现象,表现为伸长或有新叶露出,砧木也有腋芽萌发,而对照培养基中,一般在两周左右才出现生长的表象。因此,适于长寿花微嫁接的最佳暗培养时间是将嫁接苗暗培养10d。
蔗糖浓度为0g/L时,微嫁接的成活率只有3.3%;蔗糖浓度为20g/L时,成活率为70%:当蔗糖浓度为30g/L时,微嫁接的成活率最高,为96.7%:为40g/L时,微嫁接苗的成活率下降为83.3%。微嫁接苗的成活率随着蔗糖浓度的增加先提高,后下降。因此。在长寿花试管苗嫁接时应以标准的30g/L蔗糖浓度为佳。
pH值5.0-5.8时,成活率提高,当pH值为5.8时,成活率最高,为93.3%;pH值5.8-6.5时,成活率下降。因此,长寿花试管苗嫁接的最适宜pH值为5.8。
卡拉胶浓度设置为5.6g/L、5.8 g/L、6.0 g/L、6.2 g/L、6.4 g/L、6.6 g/L、6.8 g/L七个水平,结果表明:当浓度小于5.8 g/L时,接穗可以生长,但砧木出现畸形、玻璃化现象,从而失去了嫁接苗的价值,当浓度大于6.4 g/L时,嫁接苗出现了不同程度的黄化、萎蔫,不利于嫁接苗的生长。当浓度为6.0 g/L时,嫁接苗的成活率最高,达90%,且生长状态良好。因此,培养基中卡拉胶的最佳浓度为6.0 g/L。
分别在15℃、20℃、25℃、30℃条件下进行培养,结果表明:最适培养温度为25℃,成活率达到93.3%。
以牛皮纸盖、双层封口膜和塑料盖封口,结果表明:牛皮纸盖条件下,微嫁接的成活率最低,为43.3%;当用双层封口膜封口时,微嫁接成活率最高,达93.3%,并且没有污染现象:而用塑料盖封口时,成活率却只有50%并有少量的污染,因此,以双层封口膜封口来封口,对长寿花微嫁接苗的培养最为合适。
嫁接时,砧木和接穗均带叶片,有利于嫁接成活率的提高,成活率为73.3%;而砧木和接穗的长度在一定范围内对嫁接的成活影响不明显,最高为86.7%。
比较MS、1/2MS及在其中加入不同浓度的IBA、NAA的培养基中长寿花微嫁接苗的生根情况,结果表明:在1/2MS+IBA0.1mg/L+NAA0.1mg/L的生根培养基中生根率最高,为83.3%,平均根长为2.8cm且根形粗壮。
将30株生根后的嫁接苗移栽至泥炭:园土:珍珠岩=4:4:2的培养土中,20d后移栽苗的成活率为83.3%。
Kalanchoe blossfeldiana V.Poelln.are Crassulaceae.Kalanchoe Adans.perennial succulent of flowers and foliage plants..It occupys an important position on potted flowers and becomes one of the fastest-growing flower in international flower market.K.blossfeldiana V.Poelln.research so far is mainly on cultivation,propagation techniques and organizational aspects of culture in China.In foreign countries,most of the species remain in the types of physiological factors and contents of research,but the there is not any reports of tube micrografting.The species we have can not satisfy people's request,and micrografting is an utility way.At present,there is no report about effects to tube micrografting of K.blossfeldiana V.Poelln.
In this experiment,I choosed C3(violet plentiful)and C6(unknown)of six K.blossfeldiana V. Poelln.varieties(C1～C6)which leaves form and flowers color are different from each other,C3 for the scion and C6 for stock.Useing the established sterile implant system to research with grafting method,cultivation conditions,deal with stock and scion and etc.And results were as the following:
By using the establishment of a sterile implant system,we can breed fast,but unsuitable for stocks.Put C6 regenerate plants in MS + GA0.5mg/L + BA1.0mg/L culture medium for 30d,we get the adventitious buds with section between the elongation be uniform,average stem diameter increased to 0.179cm,and average height is 7.6 cm that could be suitable for the required stocks.
Using cleft grafting and top grafting method,and the result sign:the top grafting method's survival rate is 73.3%,but the cleft grafting method's survival rate is only 33.3%.Top grafting method is the best.
Put different density hormone in MS medium and the result sign:IAA is disadvantage to the plantlets.And MS+BA1.0mg/L+NAA0.1mg/L is the best medium for increasing the survival rate of grafted plantlets and its rapid growth.The survival rate is 100%,average height is 1.1cm, average growth rate is 0.009cm/d.
Cultivate the plantlets in dark for 10d,15d and 20d.The result sign:dark for 20d,survival rate is 0%;dark for 10d,survival rate is only 63.3%;both light cultivation and dark for 10d,survival rate are 100%,but the plantlets growth fast with dark for 10d.
When pH is 5.0 to 5.8,the survival rate is sublime and when pH is 5.8 to 6.5,the survival rate is breakdown.And the survival rate is highest 93.3%when pH is 5.8.
To set up density of carrageenin with 5.6g/L,5.8g/L,6.0g/L,6.2g/L,6.4g/L,6.6g/L and 6.8 g/L.Plantlets is deformed and missing the value when the density is less than 5.8g/L.When it is greater than 6.4g/L,plantlets is chlorisis and wilting.The survival rate is highest 90%and plantlets growth good with density is 6.0g/L,so 6.0g/L is the best density of carrageenin.
Put the plantlets in 15℃,20℃,25℃and 30℃.And 25℃is the optimum temperature with survival rate to be 93.3%.
Double seeling membrane is better for plantlets' growing and survival rate is 93.3%.
For improving micrografting survival rate,both of scions and rootstock should keep leaves, and survival rate is 73.3%.The effects of scions and stocks' length to survival rate would be not obvious,and the highest survival rate is 86.7%.
The optimum rooting medium is 1/2MS+ IBA0.1mg/L +NAA0.1mg/L for micrografting plantlets of K.blossfeldiana V.Poelln.,the rooting rate is 83.3%and growing strong with 2.8cm long.
Transplant 30 grafting plantlets to culture soil with Peat:Garden soil:Perlite = 4:4:2.After 20d,the survival rate is 83.3%.
本试验从六个长寿花品种中(C1～C6)选择了叶片形态与花色差异较大的两个长寿花品种C3(紫缤)和C6(未知)作为试验材料,以C3为接穗,C6为砧木。利用已经建立的尢菌体系,从嫁接方法、培养条件、砧木接穗的处理等方面对微嫁接技术体系进行了系统研究,主要研究结果如下:
利用原有的长寿花无菌快繁体系能达到快速高效的繁殖目的,但不适宜作为砧木使用。将品种C6的再生植株在MS+GA0.5mg/L+BA1.0mg/L的培养基中培养30d,得到节间伸长均匀,且平均茎粗为0.179cm,平均株高7.6cm的不定芽,适宜作为微嫁接的砧木。
嫁接方法采用劈接法和顶接法,结果表明:顶接法的嫁接成活率为73.3%,而劈接法的成活率仅为33.3%,因此,认为长寿花试管微嫁接的适宜方法为顶接法。
激素对长寿花试管微嫁接苗成活率的影响方面,研究了在MS培养基中添加不同浓度的BA、IAA、NAA,结果表明:加入IAA的培养基中嫁接苗的成活率为0,说明IAA不利于嫁接苗的成活。在培养基MS+BA1.0mg/L+NAA0.1mg/L中,嫁接的成活率为100%,平均株高1.1cm,生长速率最快为0.009cm/d,嫁接苗的接穗和砧木生长状态良好,基本上没有黄叶和落叶的现象,生长均匀,生长速率较快,茎段的粗度有所增加。因此,适于长寿花嫁接苗快速生长的最佳培养基为MS+BA1.0mg/L+NAA0.1mg/L。
暗培养10d、15d、20d与对照(光培养)相比较的结果:暗培养20d的成活率为0.15d的成活率仅63.3%,而光培养和暗培养10d的成活率达到100%,且暗培养10d时,大部分接穗都有生长的现象,表现为伸长或有新叶露出,砧木也有腋芽萌发,而对照培养基中,一般在两周左右才出现生长的表象。因此,适于长寿花微嫁接的最佳暗培养时间是将嫁接苗暗培养10d。
蔗糖浓度为0g/L时,微嫁接的成活率只有3.3%;蔗糖浓度为20g/L时,成活率为70%:当蔗糖浓度为30g/L时,微嫁接的成活率最高,为96.7%:为40g/L时,微嫁接苗的成活率下降为83.3%。微嫁接苗的成活率随着蔗糖浓度的增加先提高,后下降。因此。在长寿花试管苗嫁接时应以标准的30g/L蔗糖浓度为佳。
pH值5.0-5.8时,成活率提高,当pH值为5.8时,成活率最高,为93.3%;pH值5.8-6.5时,成活率下降。因此,长寿花试管苗嫁接的最适宜pH值为5.8。
卡拉胶浓度设置为5.6g/L、5.8 g/L、6.0 g/L、6.2 g/L、6.4 g/L、6.6 g/L、6.8 g/L七个水平,结果表明:当浓度小于5.8 g/L时,接穗可以生长,但砧木出现畸形、玻璃化现象,从而失去了嫁接苗的价值,当浓度大于6.4 g/L时,嫁接苗出现了不同程度的黄化、萎蔫,不利于嫁接苗的生长。当浓度为6.0 g/L时,嫁接苗的成活率最高,达90%,且生长状态良好。因此,培养基中卡拉胶的最佳浓度为6.0 g/L。
分别在15℃、20℃、25℃、30℃条件下进行培养,结果表明:最适培养温度为25℃,成活率达到93.3%。
以牛皮纸盖、双层封口膜和塑料盖封口,结果表明:牛皮纸盖条件下,微嫁接的成活率最低,为43.3%;当用双层封口膜封口时,微嫁接成活率最高,达93.3%,并且没有污染现象:而用塑料盖封口时,成活率却只有50%并有少量的污染,因此,以双层封口膜封口来封口,对长寿花微嫁接苗的培养最为合适。
嫁接时,砧木和接穗均带叶片,有利于嫁接成活率的提高,成活率为73.3%;而砧木和接穗的长度在一定范围内对嫁接的成活影响不明显,最高为86.7%。
比较MS、1/2MS及在其中加入不同浓度的IBA、NAA的培养基中长寿花微嫁接苗的生根情况,结果表明:在1/2MS+IBA0.1mg/L+NAA0.1mg/L的生根培养基中生根率最高,为83.3%,平均根长为2.8cm且根形粗壮。
将30株生根后的嫁接苗移栽至泥炭:园土:珍珠岩=4:4:2的培养土中,20d后移栽苗的成活率为83.3%。
Kalanchoe blossfeldiana V.Poelln.are Crassulaceae.Kalanchoe Adans.perennial succulent of flowers and foliage plants..It occupys an important position on potted flowers and becomes one of the fastest-growing flower in international flower market.K.blossfeldiana V.Poelln.research so far is mainly on cultivation,propagation techniques and organizational aspects of culture in China.In foreign countries,most of the species remain in the types of physiological factors and contents of research,but the there is not any reports of tube micrografting.The species we have can not satisfy people's request,and micrografting is an utility way.At present,there is no report about effects to tube micrografting of K.blossfeldiana V.Poelln.
In this experiment,I choosed C3(violet plentiful)and C6(unknown)of six K.blossfeldiana V. Poelln.varieties(C1～C6)which leaves form and flowers color are different from each other,C3 for the scion and C6 for stock.Useing the established sterile implant system to research with grafting method,cultivation conditions,deal with stock and scion and etc.And results were as the following:
By using the establishment of a sterile implant system,we can breed fast,but unsuitable for stocks.Put C6 regenerate plants in MS + GA0.5mg/L + BA1.0mg/L culture medium for 30d,we get the adventitious buds with section between the elongation be uniform,average stem diameter increased to 0.179cm,and average height is 7.6 cm that could be suitable for the required stocks.
Using cleft grafting and top grafting method,and the result sign:the top grafting method's survival rate is 73.3%,but the cleft grafting method's survival rate is only 33.3%.Top grafting method is the best.
Put different density hormone in MS medium and the result sign:IAA is disadvantage to the plantlets.And MS+BA1.0mg/L+NAA0.1mg/L is the best medium for increasing the survival rate of grafted plantlets and its rapid growth.The survival rate is 100%,average height is 1.1cm, average growth rate is 0.009cm/d.
Cultivate the plantlets in dark for 10d,15d and 20d.The result sign:dark for 20d,survival rate is 0%;dark for 10d,survival rate is only 63.3%;both light cultivation and dark for 10d,survival rate are 100%,but the plantlets growth fast with dark for 10d.
When pH is 5.0 to 5.8,the survival rate is sublime and when pH is 5.8 to 6.5,the survival rate is breakdown.And the survival rate is highest 93.3%when pH is 5.8.
To set up density of carrageenin with 5.6g/L,5.8g/L,6.0g/L,6.2g/L,6.4g/L,6.6g/L and 6.8 g/L.Plantlets is deformed and missing the value when the density is less than 5.8g/L.When it is greater than 6.4g/L,plantlets is chlorisis and wilting.The survival rate is highest 90%and plantlets growth good with density is 6.0g/L,so 6.0g/L is the best density of carrageenin.
Put the plantlets in 15℃,20℃,25℃and 30℃.And 25℃is the optimum temperature with survival rate to be 93.3%.
Double seeling membrane is better for plantlets' growing and survival rate is 93.3%.
For improving micrografting survival rate,both of scions and rootstock should keep leaves, and survival rate is 73.3%.The effects of scions and stocks' length to survival rate would be not obvious,and the highest survival rate is 86.7%.
The optimum rooting medium is 1/2MS+ IBA0.1mg/L +NAA0.1mg/L for micrografting plantlets of K.blossfeldiana V.Poelln.,the rooting rate is 83.3%and growing strong with 2.8cm long.
Transplant 30 grafting plantlets to culture soil with Peat:Garden soil:Perlite = 4:4:2.After 20d,the survival rate is 83.3%.
引文
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