设施百合品种筛选与“叶烧病”发生机理的研究
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摘要
百合(Lilium spp.)花型美丽,寓意美好,深得人们的喜爱。然而在北方低温弱光生长条件下,不同品种的百合栽培适应能力差异很大,且在种植5-7周(快速生长期)易发生叶烧(Upper leaf necroses),其成为影响切花和盆花质量的主要因素之一,但是其发生机理,尚未得到统一的说法。因此,在品种评价的基础上,筛选出东方百合杂种系中对叶烧病敏感程度不同的3个品种,进行不同环境条件的处理,比较不同品种发生状况和生理指标等差异,并进行相关性分析,确定叶烧病的发生机理。在此基础上,结合分子水平进行深入验证叶烧病发生机理。最后通过不同的施肥处理,对叶烧病起到有效的防治作用。该研究对百合叶烧机理的发生和防治具有重要的理论和实际应用价值。具体研究结果如下：
1.在设施栽培低温和弱光条件下,分别建立了东方百合杂种系、亚洲杂种系和OT系列百合品种的的综合评价指标。综合评价了16个东方系列百合品种,筛选出综合性状好的品种为：'Legend'、Siberia'、'Crystal blanca'、Montezuma'和‘Sorbonne'和'Red empire';综合评价了13个亚洲系列百合品种,筛选出耐低温弱光的品种为：'Prato'、'Tresor'、'Renoir'、'Elite'和'Detroit'品种；综合评价12个OT系列百合品种,筛选出耐低温弱光的品种有'Robina'、'Cocossa'、'Yelloween'、Rexona'和'Conca D.Or'。
2.选择对叶烧敏感程度不同的3个东方百合品种'Sorbonne'(易感)、'Montezuma'(中抗)和‘Siberia'(抗),设置了3种环境条件(S1-S3),在种植后4-9周,比较不同品种的形态、发病状况、生理指标和光合指标的差异,探索叶烧发生机理。结果表明在温度和湿度剧烈变化的变温环境条件下(S3),3个百合品种均显著降低了株高、叶面积、茎生根的长度、干物质分配的量和不同部位的钙含量,叶烧指数显著增加；同时3个百合品种均显著提高了丙二醛、可溶性糖、可溶性蛋白、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性,减缓了叶片叶绿素含量的增加,降低了茎生根的根系活力。通过比较结果表明：在温度和湿度剧烈变化的环境条件下,在生理水平上造成的胁迫影响大小的顺序为'Sorbonne'＞'Montezuma'＞'Siberia'。
通过种植后第7周,3个不同百合品种叶烧病指数与部分生理指标的相关性分析,结果表明：叶烧病指数与根系活力、净光合速率(Pn)和水分利用率(WUE)呈极显著负相关(P<0.01),与叶绿素(a+b)成极显著正相关。另外,3个东方百合品种的叶烧病严重指数与胞间CO2浓度(Ci)和蒸腾速率(Tr)呈正显著相关(P<0.05)；与丙二醛(MDA),气孔导度(Gs),最大潜在光化学效率(Fv/Fm)和光能转化率(ΦPsⅡ)的关系不显著。因此,根系活力、叶绿素(a+b)含量、Pn和WUE这四个生理指标可作为快速筛选百合耐叶烧病的生理指标。
3.对'Sorbonne'和‘Siberia'进行HgCl2胁迫处理,探究了叶烧病发生状况与水通道蛋自的关系。3种浓度HgCl2的处理下,'Sorbonne'和'Siberia'的部分生理指标受到不同程度的抑制,其中生长指标、根系生长、净光合速率、蒸腾速率、水分利用效率都有所降低,并且显著加重了叶烧病指数。分析出水通道蛋白在百合水分运输过程中的重要作用,推测叶烧病的发生与植物水通道蛋白有密切关系。
4.通过对东方百合的不同的施肥处理,筛选出硝酸钙(0.2mM)与微生物菌肥(0.3%)的复合肥料对叶烧发生起到有效地防治作用；通过不同基质及不同浓度的生根剂处理,筛选出适宜冬季在口光温室中栽植的'Sorbonne'|的基质为园土：草炭：牛粪：秸秆=4：1：1：1,以及浓度为‘真根’5.0ml·L-1的生根剂处理,有效地减少百合叶烧病的发生率,为百合生产提供理论依据。
Lily (Lilium spp.) is pepoles'favorite flower because of its beautiful types of flower ind its wonderful meanings. However, different lily cultivars have different adaptabilities, including upper leaf necroses (ULN) after planting5-7weeks (rapid growth period) in the low temperature and weak light conditions in the north. The ULN of lilies has already become one of the main implications of the quality of cut flowers and potted flowers of lily all over the world at present. But the disease mechanism hasn't been unified view. So, three different ULN sensitive oriental lily hybrids cultivars were screened in the different environmental conditions. Different ULN indexes and physiological indicators in the three lily cultivars were compared and associated analyzed to explore the mechanism of ULN. On this basis, the aquaporin was combined to verify the mechanism of ULN. Finally, different fertilizations were treated on lilies and effectively prevented ULN. It was important for the theoretical and practical value of the occurrence and control of ULN in lilies. The results were as follows:
1. It was built for the10screening indexes for oriental hybrids lilies, Asian lilies and'OT' hybrid lilies in the facility planting in the low temperature and weak light condition. According to the ten indexes, sixteen oriental hybrids lilies cultivars were evaluated. The research showed that'Legend','Siberia','Crystal Blanca','Montezuma'and'Sorbonne' and'Red empire'had been selected as tolerent low temperature and weak light lily cultivars. thirteen Asian lilies cultivars were screened. The result showed that'Prato','Tresor','Renoir','Elite'and'Detroit'had been selected as tolerent low temperature and weak light lily cultivars. Twelve'OT'hybrid lilies cultivars were evaluated. The research showed that 'Cocossa','Yelloween','Rexona'and'Conca D.Or' had been chosed as facility Lily Cultivars.
2. Three different ULN sensitive oriental lily hybrids cultivars, including Sorbonne'(susceptible),'Montezuma'(moderate resistant) and'the Siberia'(resistant) were screened in the three environmental conditions (S1-S3). Their morphological indexes, the ULN index, physiological indexes and photosynthetic indexes were comparised in4-9weeks after planting to explore the mechanism of ULN. The result showed that the height, leaves area, stem-root length, the distribution of dry matter and the content of calcium in different leaves parts of lilies were decreased significantly in the condition which temperature and humidity changed dramatically (S3). And in the S3, the ULN index of lilies was obviously increased. At the same time, the MDA, soluble sugar, soluble protein, enzyme activity of SOD, CAT and GR of three lily cultivars were significantly increased. In addional, the content of leaf chlorophyll increases slowly, and the stem-root activity reduced. The result showed that environmental conditions which changed dramatically in the temperature and humidity had stressed on the three oriental lily cultivars in the physiological, with different levels'Sorbonne'＞'Montezuma'＞'Siberia'.
The correlation analysis result showed that the ULN index of three lily cultivars was very significantly negatively correlated with root activity, chlorophyll (a+b), net photosynthesis (Pn) and water use efficiency (WUE)(P＜0.01), and was significantly negatively correlated with transpiration(Tr) and significantly positively correlated with transpiration intercellular concentration of carbonoxide (Ci)(P＜0.05). The ULN index of three lily cultivars was not significantly correlated with Malondialdehyde (MDA), stomatal conductance (Gs), Chlorophyll fluorescence of photosynthetic system Ⅱ (ΦPs Ⅱ) and photochemical efficiency of PS Ⅱ (Fv/Fm). According to the result, root activity, chlorophyll (a+b), Pn and WUE could become rapid screening indicators to evaluate ULN resistances among different lily cultivars in greenhouse in winter.
3. In order to explore the close relationship between the UNL index and aquaporin, it was set different concentration of HgCl2stress on different ULN sensitivity of lilies, including'Sorbonne'and'Siberia'. Their morphological indexes, root growth, the ULN index, physiological indexes and photosynthetic indexes were comparised in4-9weeks after planting to explore the mechanism of ULN. The result showed that the height, leaves area, stem-length, Pn, Tr and WUE were decreased significantly and the ULN index of lilies was obviously increased in the three concentrations of HgCl2. The homologous cloning method was used. PIP1gene of water channel proteins had been cloned by the lily leaves. The importance of the aquaporin was analysed in the water transporting process. The direct relationship of the ULN index and plant aquaporin was found.
4. Different fertilizations were treated on oriental lily hybrids. The calcium nitrate (0.2mM) with the biological fertilizer (0.3%) was screened to prevent effectively the ULN. Through different concentrations of rooting powder and media were used. The most suitable concentration was selected. The most suitable media and root-growing paste for 'Sorbonne'which are cultivated in greenhouse are soil.grass carbon,cow dropping,straw (4:1:1:1) and "Zhen gen"(5.0ml L-1). The research provided a theoretical basis for the production of lilies.
1.在设施栽培低温和弱光条件下,分别建立了东方百合杂种系、亚洲杂种系和OT系列百合品种的的综合评价指标。综合评价了16个东方系列百合品种,筛选出综合性状好的品种为：'Legend'、Siberia'、'Crystal blanca'、Montezuma'和‘Sorbonne'和'Red empire';综合评价了13个亚洲系列百合品种,筛选出耐低温弱光的品种为：'Prato'、'Tresor'、'Renoir'、'Elite'和'Detroit'品种；综合评价12个OT系列百合品种,筛选出耐低温弱光的品种有'Robina'、'Cocossa'、'Yelloween'、Rexona'和'Conca D.Or'。
2.选择对叶烧敏感程度不同的3个东方百合品种'Sorbonne'(易感)、'Montezuma'(中抗)和‘Siberia'(抗),设置了3种环境条件(S1-S3),在种植后4-9周,比较不同品种的形态、发病状况、生理指标和光合指标的差异,探索叶烧发生机理。结果表明在温度和湿度剧烈变化的变温环境条件下(S3),3个百合品种均显著降低了株高、叶面积、茎生根的长度、干物质分配的量和不同部位的钙含量,叶烧指数显著增加；同时3个百合品种均显著提高了丙二醛、可溶性糖、可溶性蛋白、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽还原酶(GR)活性,减缓了叶片叶绿素含量的增加,降低了茎生根的根系活力。通过比较结果表明：在温度和湿度剧烈变化的环境条件下,在生理水平上造成的胁迫影响大小的顺序为'Sorbonne'＞'Montezuma'＞'Siberia'。
通过种植后第7周,3个不同百合品种叶烧病指数与部分生理指标的相关性分析,结果表明：叶烧病指数与根系活力、净光合速率(Pn)和水分利用率(WUE)呈极显著负相关(P<0.01),与叶绿素(a+b)成极显著正相关。另外,3个东方百合品种的叶烧病严重指数与胞间CO2浓度(Ci)和蒸腾速率(Tr)呈正显著相关(P<0.05)；与丙二醛(MDA),气孔导度(Gs),最大潜在光化学效率(Fv/Fm)和光能转化率(ΦPsⅡ)的关系不显著。因此,根系活力、叶绿素(a+b)含量、Pn和WUE这四个生理指标可作为快速筛选百合耐叶烧病的生理指标。
3.对'Sorbonne'和‘Siberia'进行HgCl2胁迫处理,探究了叶烧病发生状况与水通道蛋自的关系。3种浓度HgCl2的处理下,'Sorbonne'和'Siberia'的部分生理指标受到不同程度的抑制,其中生长指标、根系生长、净光合速率、蒸腾速率、水分利用效率都有所降低,并且显著加重了叶烧病指数。分析出水通道蛋白在百合水分运输过程中的重要作用,推测叶烧病的发生与植物水通道蛋白有密切关系。
4.通过对东方百合的不同的施肥处理,筛选出硝酸钙(0.2mM)与微生物菌肥(0.3%)的复合肥料对叶烧发生起到有效地防治作用；通过不同基质及不同浓度的生根剂处理,筛选出适宜冬季在口光温室中栽植的'Sorbonne'|的基质为园土：草炭：牛粪：秸秆=4：1：1：1,以及浓度为‘真根’5.0ml·L-1的生根剂处理,有效地减少百合叶烧病的发生率,为百合生产提供理论依据。
Lily (Lilium spp.) is pepoles'favorite flower because of its beautiful types of flower ind its wonderful meanings. However, different lily cultivars have different adaptabilities, including upper leaf necroses (ULN) after planting5-7weeks (rapid growth period) in the low temperature and weak light conditions in the north. The ULN of lilies has already become one of the main implications of the quality of cut flowers and potted flowers of lily all over the world at present. But the disease mechanism hasn't been unified view. So, three different ULN sensitive oriental lily hybrids cultivars were screened in the different environmental conditions. Different ULN indexes and physiological indicators in the three lily cultivars were compared and associated analyzed to explore the mechanism of ULN. On this basis, the aquaporin was combined to verify the mechanism of ULN. Finally, different fertilizations were treated on lilies and effectively prevented ULN. It was important for the theoretical and practical value of the occurrence and control of ULN in lilies. The results were as follows:
1. It was built for the10screening indexes for oriental hybrids lilies, Asian lilies and'OT' hybrid lilies in the facility planting in the low temperature and weak light condition. According to the ten indexes, sixteen oriental hybrids lilies cultivars were evaluated. The research showed that'Legend','Siberia','Crystal Blanca','Montezuma'and'Sorbonne' and'Red empire'had been selected as tolerent low temperature and weak light lily cultivars. thirteen Asian lilies cultivars were screened. The result showed that'Prato','Tresor','Renoir','Elite'and'Detroit'had been selected as tolerent low temperature and weak light lily cultivars. Twelve'OT'hybrid lilies cultivars were evaluated. The research showed that 'Cocossa','Yelloween','Rexona'and'Conca D.Or' had been chosed as facility Lily Cultivars.
2. Three different ULN sensitive oriental lily hybrids cultivars, including Sorbonne'(susceptible),'Montezuma'(moderate resistant) and'the Siberia'(resistant) were screened in the three environmental conditions (S1-S3). Their morphological indexes, the ULN index, physiological indexes and photosynthetic indexes were comparised in4-9weeks after planting to explore the mechanism of ULN. The result showed that the height, leaves area, stem-root length, the distribution of dry matter and the content of calcium in different leaves parts of lilies were decreased significantly in the condition which temperature and humidity changed dramatically (S3). And in the S3, the ULN index of lilies was obviously increased. At the same time, the MDA, soluble sugar, soluble protein, enzyme activity of SOD, CAT and GR of three lily cultivars were significantly increased. In addional, the content of leaf chlorophyll increases slowly, and the stem-root activity reduced. The result showed that environmental conditions which changed dramatically in the temperature and humidity had stressed on the three oriental lily cultivars in the physiological, with different levels'Sorbonne'＞'Montezuma'＞'Siberia'.
The correlation analysis result showed that the ULN index of three lily cultivars was very significantly negatively correlated with root activity, chlorophyll (a+b), net photosynthesis (Pn) and water use efficiency (WUE)(P＜0.01), and was significantly negatively correlated with transpiration(Tr) and significantly positively correlated with transpiration intercellular concentration of carbonoxide (Ci)(P＜0.05). The ULN index of three lily cultivars was not significantly correlated with Malondialdehyde (MDA), stomatal conductance (Gs), Chlorophyll fluorescence of photosynthetic system Ⅱ (ΦPs Ⅱ) and photochemical efficiency of PS Ⅱ (Fv/Fm). According to the result, root activity, chlorophyll (a+b), Pn and WUE could become rapid screening indicators to evaluate ULN resistances among different lily cultivars in greenhouse in winter.
3. In order to explore the close relationship between the UNL index and aquaporin, it was set different concentration of HgCl2stress on different ULN sensitivity of lilies, including'Sorbonne'and'Siberia'. Their morphological indexes, root growth, the ULN index, physiological indexes and photosynthetic indexes were comparised in4-9weeks after planting to explore the mechanism of ULN. The result showed that the height, leaves area, stem-length, Pn, Tr and WUE were decreased significantly and the ULN index of lilies was obviously increased in the three concentrations of HgCl2. The homologous cloning method was used. PIP1gene of water channel proteins had been cloned by the lily leaves. The importance of the aquaporin was analysed in the water transporting process. The direct relationship of the ULN index and plant aquaporin was found.
4. Different fertilizations were treated on oriental lily hybrids. The calcium nitrate (0.2mM) with the biological fertilizer (0.3%) was screened to prevent effectively the ULN. Through different concentrations of rooting powder and media were used. The most suitable concentration was selected. The most suitable media and root-growing paste for 'Sorbonne'which are cultivated in greenhouse are soil.grass carbon,cow dropping,straw (4:1:1:1) and "Zhen gen"(5.0ml L-1). The research provided a theoretical basis for the production of lilies.
引文
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