Ice‐‐bacteria as antagonists in biological frost protection

Investigated was the efficacy of 4 ice nucleation‐inactive (ice^‐) bacterial strains (A506, GSPB 1147, 1181, 2357) at 4 ice nucleation active (ice⁺) strains (553, 554, GSPB 1139, 2035) on the surface of 4 culture crops (corn, tomato, bush‐ and field bean). Examinated was the reduction of frost damage at the used culture crops by inoculation with ice⁺ and ice^‐ strains at 4 different variants (A‐D) under laboratory conditions. The ice nucleation activity was determined by the tube‐freezing‐assay. An effective reduction of ice⁺ bacteria was possible, when plant surfaces were preinoculated (3days) with ice^‐bacteria before ice⁺ bacterial strains colonized the surfaces of plants. A statistical comparison of mean values of obtained results showed, the ice crystallization (INT) by inoculation started at ‐3°C and in mean (MNT) below ‐5°C independent of the used ice⁺ and ice^‐ strains. Obtained differences were not significant. As untreated as colonized plants with antagonists started to freeze at ‐5 and ‐6°C and in average at ‐7°C. However, the preinoculation resulted in 1.38 K differences for the temperatures, at which 50% of leaves were frozen (PROZ 50) in favour of preinoculated plants. This difference in freeze temperatures by preinoculation with ice^‐ bacteria was discussed as a possible method of frost protection.     

菌种保存方法对冰核细菌冰核活性的影响

本文测定了5种保存方法及3种保存温度,对4株代表3个属4个种(变种)的冰核细菌冰棱活性的影响。结果表明,不同保存方法对冰核细菌的存活力和冰核活性存在着不同程度的影响,保存温度越高,对冰核活性的影响越大,不同菌种对保存的敏感性不同。所适合的保存方法也不同。真空冷冻干燥和灭菌水于-20℃冷冻保存对各菌株的存活力和冰核活性的影响都较小,适用于一般冰棱细菌的长期保存。而10%甘油冷冻保存则不适用于冰核细菌的保存。     

冰核真菌的冰核活性及其种类鉴定*

作者从1993年起至今,从已分离和搜集到的500多株真菌中筛选出在-5℃具有冰核活性的真菌6株,它们-5℃的冻滴率高低顺序为F9502（100%）=F9501（100%）＞F9401（96％）=AS3．494（96%）＞F9801（94%）＞F9802（92%）,结冰点高低顺序为F9502（-2．7℃）＞F9501（-3．4℃）＞F9401（-4.4℃）＞AS34594（-4.5℃）＞F9801（-4．7℃）＞F9402（-4．8℃）,以F9502菌株活性强而稳定。经鉴定确定：F9401和F9402菌株为FusartumSportFichioldesSherb．;AS3．4594菌株为FavenaceumSacc．;F9501和F9502菌株为F.gramlnearumSchwabe;F9801菌株为F．monilijormeSheldon。F．Sportrlchloides和F．graminearum作为冰核真菌未见报道。本结果为揭示冰核真菌与植物冻害关系及冰核真菌开发应用研究提供了菌种资源,有重要应用价值。     

冰核真菌的冰核活性及其种类鉴定

作者从1993年起至今,从已分离和搜集到的500多株真菌中筛选出在-5℃具有冰核活性的真菌6株,它们-5℃的冻滴率高低顺序为F9502（100%）=F9501（100%）＞F9401（96％）=AS3．494（96%）＞F9801（94%）＞F9802（92%）,结冰点高低顺序为F9502（-2．7℃）＞F9501（-3．4℃）＞F9401（-4.4℃）＞AS34594（-4.5℃）＞F9801（-4．7℃）＞F9402（-4．8℃）,以F9502菌株活性强而稳定。经鉴定确定：F9401和F9402菌株为FusartumSportFichioldesSherb．;AS3．4594菌株为FavenaceumSacc．;F9501和F9502菌株为F.gramlnearumSchwabe;F9801菌株为F．monilijormeSheldon。F．Sportrlchloides和F．graminearum作为冰核真菌未见报道。本结果为揭示冰核真菌与植物冻害关系及冰核真菌开发应用研究提供了菌种资源,有重要应用价值。     

冰核细菌表达冰核蛋白特性的研究

选用１００２５Ａ和ＱＦ－９５－Ｆ１９两株分离自杨树的冰核活性细菌,探讨了两株菌不同生长阶段与它们冰核活性表达的特性。实验结果显示,冰核活性细菌在ＭＰＤＡ培养液中表达冰核蛋白的特性及活性与细菌浓度、菌龄以及培养的环境条件相关,两株菌在表达冰核活性时对培养基的营养组分没有表现出特殊的要求。同时还进一步阐明了不同生长温度冰核活性细菌对冰核蛋白表达的影响。     

光化学反射指数在比较葡萄叶片耐霜冻能力中的应用

以12个葡萄(Vitis vinifera)品种为试材,在秋季采集叶片,进行霜冻处理,测定了霜冻过程中叶片光化学反射指数(PRI)的变化。提取有效参数,采用隶属函数法比较了不同叶片的耐霜冻能力。结果表明,随着温度的降低,叶片光化学反射指数呈下降趋势。在–8–0°C范围内,PRI及其相对值Rt与温度都存在显著的线性关系;叶片PRI在0°C、–8°C时的相对值R0、R–8,以及–8–0°C范围中,Rt与温度的回归直线斜率SLP都具有品种特异性,可以反映不同品种的耐霜冻能力。隶属函数法结合聚类分析结果显示,在12个候选品种中,抗霜冻能力最好的是Cabernet Sauvignon,其R0、R–8和SLP分别为72.3%、50.19%和2.78。     

杉木人工林冰雪灾害受灾和萌生情况调查

以九龙山国家级自然保护区的主要公益林杉木人工林作为研究对象,选择具有代表性的受灾区域,自海拔700 m至900 m设置样地,共10块样地,总面积4 000 m2。分析其受2008年初严重冰雪灾害破坏及灾后萌生情况与胸径、树高、尖削度、海拔之间的关系。结果表明：1）杉木人工林损害严重,其中断冠、断干植株比例较高,而掘根和冻死植株相对较少;杉木具有极强的萌生能力,以断冠杉木的萌生能力最强。2）不同胸径、树高、尖削度的杉木对于冰雪灾害的抵御能力及萌生能力存在一定差异,小径级矮小的个体受灾较重,易被冻死和掘根,大径级的较高个体受灾相对较轻,主要表现为断冠和断干危害,且不同径级下的受灾杉木上部萌生均占较大比例。3）杉木的抵御能力和萌生能力与海拔存在一定的关系,高海拔地区杉木受灾较重,而中海拔地区的杉木萌生能力相对较强。     

水杨酸对大叶黄杨茎抗寒性和电阻抗图谱参数的影响

在抗寒锻炼前,对当年生大叶黄杨(Euonymus japonicus)扦插苗喷施不同浓度水杨酸,用电阻抗图谱(EIS)法和电导(EL)法估测茎的抗寒性,以探明抗寒锻炼期间水杨酸对大叶黄杨抗寒性和电阻抗图谱参数的影响,找到适合不经冷冻处理估测大叶黄杨茎抗寒性和经冷冻处理后测定其抗寒性的EIS参数。结果表明,水杨酸处理能够提高大叶黄杨茎的抗寒性,最适浓度为5.0mmol.L-1;不经冷冻处理茎的EIS参数电阻率r和r1、胞外电阻率re、胞内电阻率ri、弛豫时间τ、弛豫时间分布系数ψ与EL法测定的抗寒性有较高的相关性(r=0.70~0.87),说明不经冷冻处理样本用以上参数估测大叶黄杨茎抗寒性是可行的,r1为最佳参数;冷冻处理后茎的re、τ、ri求得的抗寒性与EL法测定的抗寒性有较高的相关性(r=0.85~0.94),说明冷冻处理后re、τ、ri可以作为测定大叶黄杨茎抗寒性的参数,re为最佳参数。     

Frost hardiness development in Pinus silvestris seedlings in response to fertilization

The effects of different levels of nitrogen on freezing survival, the amounts of chlorophylls and β-carotene, the ratios Chl a/Chl b, and (Chl a + Chl b)/β-carotene in hardened and unhardened seedlings of Scots Pine (Pinus silvestris L.) have been investigated. There was. no correlation between frost hardiness and nitrogen content in the unhardened seedlings. However, the hardy seedlings with the highest nitrogen content, showed a decrease in frost hardiness of 3°C. The amounts of chlorophylls and β-carotene increased for all the treatments during hardening, except for the seedlings that had the highest nitrogen content; so that the effect of fertilizer was more evident in non-hardened than in hardened plants. The ratios Chl a/Chl b decreased during hardening in all the treatments. This was an effect of a larger increase of Chl b than Chl a. The ratios of Chl a + Chl b/β-carotene also decreased during hardening except for the plants that were fertilized with the second lowest amount of nitrogen. Since the amounts of Chl a and Chl b increase during hardening the amount of β-carotene increases even more.
The growth rates of the seedlings were closely related to the nitrogen content for the different treatments. All plants were infected with ectotrophic mycorrhizae, which resulted in an enhancement of ion transportation from the soil to the plants. This is important when the supply of ions is low.     

Molecular cloning of an ice nucleation gene from Erwinia ananas and its expression in Escherichia coli

Abstract An approximately 7 kbp genomic DNA fragment was cloned from an ice nucleation-active (ina) strain of Erwinia ananas and defined as to its restriction enzyme site. When the DNA fragment was introduced into E. coli MM294, a potent ice nucleation activity was expressed. Both 0.7 kbp truncation from the 5'-end and 1.7 kbp truncation from the 3'-end were also effective in expressing the ice nucleation activity in E. coli . Therefore, the resulting DNA fragment of approximately 5 kbp was considered to be an ina gene and named ina A. It existed as a unique gene in this strain of E. ananas . No corresponding ina gene existed in an ice nucleation-inactive strain of E. milletiae .     

云南植物上冰核活性细菌鉴定

从云南植物上分离到９２株冰核活性细菌,并进行了鉴定。其中菠萝欧文氏菌６１株,占６６．３％;草生欧文氏菌２株,占２．２％;丁香假单胞菌２１株,占２２．８％;黄瓜角斑病菌２株,占２．２％;菜豆荚斑假单胞菌６株,占６．５％。云南省冰核活性细菌的优势种类是菠萝欧文氏菌,其次是丁香假单胞菌类。     

Dynamic thermal time model of cold hardiness for dormant grapevine buds

Background and Aims

Grapevine (Vitis spp.) cold hardiness varies dynamically throughout the dormant season, primarily in response to changes in temperature. The development and possible uses of a discrete-dynamic model of bud cold hardiness for three Vitis genotypes are described.

Methods

Iterative methods were used to optimize and evaluate model parameters by minimizing the root mean square error between observed and predicted bud hardiness, using up to 22 years of low-temperature exotherm data. Three grape cultivars were studied: Cabernet Sauvignon, Chardonnay (both V. vinifera) and Concord (V. labruscana). The model uses time steps of 1 d along with the measured daily mean air temperature to calculate the change in bud hardiness, which is then added to the hardiness from the previous day. Cultivar-dependent thermal time thresholds determine whether buds acclimate (gain hardiness) or deacclimate (lose hardiness).

Key Results

The parameterized model predicted bud hardiness for Cabernet Sauvignon and Chardonnay with an r² = 0·89 and for Concord with an r² = 0·82. Thermal time thresholds and (de-)acclimation rates changed between the early and late dormant season and were cultivar dependent but independent of each other. The timing of these changes was also unique for each cultivar. Concord achieved the greatest mid-winter hardiness but had the highest deacclimation rate, which resulted in rapid loss of hardiness in spring. Cabernet Sauvignon was least hardy, yet maintained its hardiness latest as a result of late transition to eco-dormancy, a high threshold temperature required to induce deacclimation and a low deacclimation rate.

Conclusions

A robust model of grapevine bud cold hardiness was developed that will aid in the anticipation of and response to potential injury from fluctuations in winter temperature and from extreme cold events. The model parameters that produce the best fit also permit insight into dynamic differences in hardiness among genotypes.     

Age-related differences in frost sensitivity of the photosynthetic apparatus of two Plagiomnium species

Shoots of two species of moss, Plagiomnium undulatum (Hedw.) Kop. and Plagiomnium affine (Funck) Kop., were subjected to freezing at various temperatures. After thawing, the activities of different photosynthetic reactions were determined in relation to the ages of the leaves. Analysis of the fast kinetics of chlorophyll-a fluorescence of individual leaves showed that young and old tissues were considerably less frost tolerant than mature ones. In principle, the pattern of freeze inactivation of photosynthetic reactions resembles that observed in higher plants. The decreases in the amplitude of F_v (variable fluorescence) and the ratio of F_v to F_m (maximum fluorescence) with increasing freezing stress reflect a progressive inactivation of photosystem II (PSII)-mediated electron transport, i.e. inhibition of photoreaction to photochemistry and-or electron donation to the photochemical reaction, and thus a decline in the potential photochemical efficiency of PSII. The insignificant change in the F₀ (constant fluorescence) level during progressive decline of F_v indicates that the excitation-energy transfer between antenna pigments and from those to reaction centres of PSII was little impaired by lethal freezing stress. Sugar analyses of various stem sections showed that ontogenetic variation in the frost tolerance of leaves cannot be attributed to differences in the cellular levels of sucrose, glucose and fructose.Abbreviations and Symbols DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_m maximum fluorescence - F₀ constant (initial) fluorescence - F_v variable fluorescence