林业部最新科技成果推广项目

林业部最新科技成果推广项目一、用材林良种及丰产栽培技术１．欧洲黑杨抗食叶害虫转基因植株１９２、１２和１５３号的推广主要内容：采用基因工程方法，将Ｂｔ杀虫毒蛋白导入欧洲黑杨植株的染色体中，获１９２、１２和１５３号３种植株，为食叶害虫泛滥地区提供了抗虫品...     

雄性毛白杨离体叶片再生及抗虫基因转化

用雄性毛白杨（Ｐｏｐｕｌｕｓｔｏｍｅｎｔｏｓａ）试管植株无菌叶片作外植体，筛选出诱导不定芽分化、增殖和生根培养基。在组培室的光照条件下培养３７ｄ，平均每片叶产生２５个不定芽。已建立雄性毛白杨最佳转化和再生系统。用含有完全改造的Ｂｔ抗虫基因表达载体ｐＢ４８．７和部分改造的Ｂｔ抗虫基因表达载体ｐＢ４８．６转化雄性毛白杨，经在含有卡那霉素的培养基上选择和ＰＣＲ检测，已获得初步确定的转基因植株。     

表达豇豆胰蛋白酶抑制剂转基因杨树的虫试试验

将转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨回交杂种 [(毛新杨×毛白杨 )×毛白杨 ]]于春季萌发前截干 .用萌条上的树叶对 3种主要杨树害虫 :天幕毛虫 (Malacosomadisstria)、舞毒蛾 (Lymant riadisparL .)和杨雪毒蛾 (StilpnotiacandidaStaudinger)进行离体虫试试验 ,年底测量各转基因株系萌条树高和地径 ,以衡量其生长性状 .结果发现 ,与对照相比 ,转基因株系的树高和地径没有显著降低 ,说明外源基因的导入没有影响转基因杨树的速生特性 ,相反 ,TG0 4的树高和地径较对照有明显的提高 ,这可能是外源导入后促进转基因杨树生长的结果 ;转基因植株均能显著提高害虫的死亡率 ,降低幼虫的排粪量、蛹重、体重增加量和成虫的产卵量 ;其中 ,TG0 7、TG0 8和TG713个转基因株系的抗虫特性较为突出 ,表明CpTI基因在它们的树体内表达较为活跃和稳定     

抗虫转基因欧美杨的培育

以携带３５Ｓ－Ω－Ｂ．ｔ－Ｎｏｓ嵌合基因的双元载体农杆菌ｐＢ４８．２１４和ｐＢ４８２．１５（Ｂ．ｔ基因片断长度分别为２．１ｋｂ和１．８ｋｂ），转化欧美杨叶片和茎段外植体，共获得２２５株转化再生植株。以舞毒蛾（ＬｙｍａｎｔｒｉａｄｉｓｐａｒＬｉｎｎａｅｕｓ）幼虫进行生物测定后，获得杀虫率分别为５５—８０％的抗虫转基因植株共３株（Ｒ－１３５，Ｒ－１２３和Ｒ－１４０），经组培苗和苗圃移栽植株的ＰＣＲ分析，苗圃移栽植株ＰＣＲ产物杂交和Ｓｏｕｔｈｅｒｎｂｌｏｔ分析表明，Ｂ．ｔ毒蛋白基因已整合到上述抗虫植株的细胞ＤＮＡ中，并表达出杀虫活性。     

转Bt基因和蛋白酶抑制剂基因杨树抗虫性的研究

虫害是杨树造林生产中所面临的一大难题,通过农杆菌介导,将经过人工改造的苏云金杆菌杀虫结晶蛋白基因转化杨树,得到杨树再生植株,然后,再将蛋白酶抑制剂基因（ＰＩ）导入已含Ｂｔ基因的转基因杨树,经ＰＣＲ检测和Ｓｏｕｔｈｅｒｎ杂交分析证明,最终获得既含有Ｂｔ基因又含有蛋白酶抑制基因的转基因杨树植株。利用这种杨种叶片饲同喂舞毒蛾幼虫的杀虫试验结果表明,转基因杨树具有明显的杀虫活性,同时表明含有Ｂｔ基因和蛋白     

转不同抗虫基因741杨的抗虫选择性

以转抗虫基因(BtCry3A)741杨7个株系和转双抗虫基因(BtCrylAc API)741杨2个株系为材料,从分子生物学检测、毒蛋白的表达、抗虫性检测3个方面,研究转不同Bt基因741杨对不同类型害虫的抗虫选择性.结果表明:2种Bt抗虫基因分别在转基因741杨不同株系中稳定存在.转BtCrylAc的2个株系毒蛋白含量分别为0.012 7%和0.009 9%,具高抗虫性的株系pb29的毒蛋白含量高于中等抗虫性的株系pb17.转BtCry3A基因的5个株系毒蛋白含量为0.067 8%～0.152 1%,不同株系间存在一定差异.BtCry3型的毒蛋白表达量非常高,是BtCry1型的10倍.用对鳞翅目害虫具高抗虫性和中等抗虫性的转双抗虫基因(BtCrylAc API)741杨株系叶片饲养鞘翅目害虫柳蓝叶甲,幼虫死亡率达到28.4%～42.8%,转基因株系与未转基因的对照没有明显差异.用转BtCry3A基因741杨不同株系叶片饲养柳兰叶甲幼虫,均表现出高抗虫性,死亡率在2天之内均可达到100%;而饲养杨扇舟蛾幼虫,死亡率为0～42.86%,未表现出明显抗虫性,与未转基因对照没有明显差异.     

双抗虫基因对三倍体毛白杨的转化和抗虫性表达

建立三倍体毛白杨叶片再生体系,用部分改造BtCry1Ac基因与慈菇蛋白酶抑制剂(API)基因构建的双抗虫基因表达载体,通过农杆菌介导法转化三倍体毛白杨无性系,在含卡那霉素50 mg·L-1的分化培养基上诱导产生不定芽的叶片占18%,在生根培养基上对转基因植株做进一步筛选,获得50个转化再生株系.PCR检测表明:80%的植株呈现阳性反应,部分株系进行Southern Blot检测,证明外源基因已经插入杨树基因组中.用Bt毒蛋白抗血清进行ELSA检测,结果表明:7个转基因株系都有Bt杀虫蛋白表达,表达量最高的株系约占叶总可溶性蛋白的0.016 1%.用经分子生物学检测的28个转基因株系叶片进行舞毒蛾和杨扇舟蛾幼虫饲虫试验,结果表明:不同株系幼虫杀死率明显不同,有39.3%的株系对舞毒蛾和杨扇舟蛾幼虫的致死率在80%以上;25.0%的株系对两种害虫的幼虫死亡率均在60%～80%之间;另有35.7%的株系幼虫死亡率在50%以下,有些株系基本未表现出抗虫性.同时具高抗虫性的转基因株系能够明显抑制存活幼虫的生长和发育.     

延缓害虫对Bt毒蛋白耐受性的育种策略

在分析害虫对转 Bt毒蛋白基因植株产生耐受性的特点、发生机制的基础上 ,重点探讨了为延缓害虫对 Bt毒蛋白耐受性的育种策略 :(1)培育双价或多价 Bt毒蛋白抗虫品种 ;(2 )寻找能使 Bt毒蛋白基因在植物中高效表达的启动子 ;(3 )使用组织特异型或诱导型启动子以减轻对害虫持续的选择压 ;(4 )将 Bt毒蛋白基因转入叶绿体基因组以解决原核与真核细胞对遗传密码子偏爱的不同 ;(5 )通过突变等技术改变 Bt毒蛋白一级结构 ,从分子水平延缓害虫产生耐受性     

表达豇豆胰蛋白酶抑制剂转基因杨树的蛋白检测

对转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨杂种 [(毛新杨×毛白杨 )×毛白杨 ]的可溶性总蛋白和胰蛋白酶抑制剂蛋白的含量进行测定 .结果发现 ,与未转基因植株相比 ,所有供试转基因株系叶片中的总可溶性蛋白含量明显增加 ,但老叶的含量高于嫩叶 ,这表明转基因株系可溶性总蛋白含量增加可能是CpTI基因表达的结果或由于外源基因导入后引起杨树基因组中某些自身基因的表达所致 .转基因株系叶片中有较高含量CpTI,而对照叶片则检测不到CpTI,这进一步证实了CpTI基因已在转基因株系中得到稳定表达 .进一步比较发现 ,在供试的 5个无性系中 ,TG0 7、TG0 4和TG71在总蛋白含量和CpTI含量增加较为明显 .另外 ,聚丙烯酰胺凝胶电泳胶分析发现 ,与未转基因植株相比 ,在所有供试转基因株系叶片中均出现一条分子量为 11.3kD的清晰蛋白带     

741杨双Bt基因的遗传转化及转基因株系的抗虫性

以含有Cry3Aa基因的重组质粒pBCC3为基础,利用PCR和DNA重组技术,从pBCC3中克隆出抗虫基因Cry3Aa,将其正向插入载体pCAMBIA1305的CaMV35S启动子和NOS终止子之间,构建成pCAMBIA1305-Cry3Aa植物表达载体,并导入根癌农杆菌EHA105。采用农杆菌介导的遗传转化法,将Cry3Aa基因转入已转Cry1Ac+API基因的741毛白杨无性系pB29中,获得转双Bt基因的741杨。在含潮霉素的培养基中进行多次继代筛选,获得抗性稳定的无性系9个,编号为pCCA1—pCCA9。采用特异引物分别对转基因植株进行PCR检测,结果显示Cry1Ac基因稳定存在于pB29中,Cry3Aa基因已整合到各无性系的基因组DNA中。ELISA毒蛋白检测,转基因株系都有Cry1Ac和Cry3Aa杀虫蛋白表达。用转基因植株叶片进行柳蓝叶甲(鞘翅目)和美国白蛾(鳞翅目)室内饲虫试验结果表明,转基因植株具有双抗性。根据对测试昆虫的致死率划分高中低3个抗性水平,其中pCCA2,pCCA5,pCCA6,pCCA9具有双高抗;pCCA3,pCCA4和pCCA7对柳蓝叶甲表现出中、低抗性,对美国白蛾则高抗;而pCCA1表现对美国白蛾的极低抗性,对柳蓝叶甲则高抗。     

Browsing Damage on Pine (Pinus sylvestris and P. contorta) by a migrating moose (Alces alces) Population in Winter: Relation to Habitat Composition and Road Barriers

In Scandinavia, moose (Alces alces L.) sometimes cause severe browsing damage to economically-important pine. Moose-vehicle accidents have spurred construction of fences along roads, and these may interfere with moose migration between summer and winter ranges, or the road alone may be a barrier. If this happens and moose build up along roads, landowners may suffer economically. Therefore, this study investigated whether roads, fences or other factors influence the use of young pine stands by moose. Eighty stands along roads in northern Sweden were evaluated in which individually-browsed branches were counted on 9972 pines. Moose browsing was not significantly related to birch (Betula pendula Roth, B. pubescens Ehrh.) density, nor did it differ between pines (Pinus contorta Douglas or P. sylvestris L.). However, increased pine density, site productivity and proximity to a highway were associated with increased browsing. Further large-scale studies are needed to understand moose habitat selection and the effects of roads.     

Effects of Root Freezing on the Physiology and Growth of Picea glauca,Picea mariana and Pinus banksiana Seedlings Under Different Soil Moisture Regimes

The root systems of 2-yr-old Picea glauca, Picea mariana and Pinus banksiana seedlings were submitted to various frost temperatures during an artificial frost to induce different levels of root damage. Frost-damaged and control seedlings were placed in a greenhouse under high and low soil moisture regimes. Seedling growth and physiology were evaluated periodically. Seedling survival was reduced when root damage reached levels of 60-80%. Root systems of all three species showed partial to total recovery by the end of the experiment. In general, root freezing damage caused reductions in seedling growth, with these reductions becoming less significant over time. Root damage had little to no effect on black spruce and jack pine seedling physiology, while white spruce CO 2 uptake decreased with increasing root damage. Shoot nitrogen content of all three species decreased slightly with increasing root damage.     

Stand Structure in Pine,Spruce and Deciduous Forests: A Comparison Between Finnish and Russian Karelia

This study investigated the stand structure in pine, spruce and deciduous forests in the border district of Finland and Russia. A total of 46 mature forest stands was selected as pairs, the members of each pair being as similar as possible with respect to their forest site type, age, moisture and topography. The stands were then compared between the two countries by means of basal areas and number of stems. The proportions of dominating tree species were 2-12% lower, and correspondingly the proportions of secondary tree species higher, in Russian forests. The density of the forest stock was also higher in each forest type in Russia. The forests in the two countries differed most radically in terms of the abundance of dead trees. The amount was two to four times higher in Russian deciduous and spruce forests, and in pine forests the difference was 10-fold. The stand structures indicated that Russian coniferous stands, in particular, were more heterogeneous than intensively managed pine and spruce stands in Finland.     

Growth and Terminal Bud Formation in Picea abies Seedlings Grown with Alternating Diurnal Temperature and Different Light Qualities

The root collar diameter and the height:diameter ratio are of particular importance in container-grown seedlings where a high density in the containers may produce spindly seedlings. Temperature regimes and light quality are known to affect plant growth. The aim of this study was to identify responses in Picea abies (L.) Karst. seedlings grown with light providing different red:far-red ratios and under temperature regimes with alternating day (DT) and night temperature (NT) from negative (DT < NT) to positive (DT > NT) difference (DIF) between DT and NT. Experiments were conducted in controlled environment chambers and in a daylight phytotron. Only limited thermoperiodic responses appeared in P . abies seedlings with respect to seedling height and dry weight accumulation. The formation of terminal buds, however, was clearly delayed in seedlings grown at negative DIF. The results indicate a requirement for day extension light that is high in far-red, to prevent terminal bud formation under natural short-day conditions. An extended study should be conducted to clarify the minimum level of light intensity and the optimal light quality needed to prevent terminal bud formation under natural short-day conditions.     

CAD与建筑设计的关系

本文分析了CAD在设计中引起的正面和负面影响,并进行了系统的阐述,从而使设计者在应用中能够保持客观的态度。     

Natural Regeneration in Clear-cuts: Effects of Scarification,Slash Removal and Clear-cut Age

The effects of soil scarification (mounding), slash removal and clear-cut age on the natural regeneration in clear-cuts was evaluated using data from four sites in southern Sweden. The treatments were carried out during a good seed and establishment year for birch ( Betula pubescens Ehrh. and B. pendula Roth). Scarification had the strongest positive effect on the density of naturally regenerated seedlings, especially in birch, but also in pine ( Pinus sylvestris L.) and spruce [ Picea abies (L.) Karst]. Slash removal had a positive effect on birch density. No statistically significant effect of clear-cut age was found. In addition, no statistically significant interactions between clear-cut age and scarification or slash removal were found. The ingrowth of field vegetation was the fastest in areas that were not scarified, less rapid in areas scarified on old clear-cuts, and the slowest after scarification in fresh clear-cuts. In conclusion, it may be possible to control the density of birch during a good establishment year for birch. If birch is desired, the best combination of treatments is to remove the slash and scarify; otherwise, these treatments should be avoided.     

Risk Perception Among Non-industrial Private Forest Owners

Based on an enquiry, risk perception among non-industrial private forest owners is described in relation to climate change and forestry hazards. Of the respondents, 11% took action to remedy the effects of climate change. Out of a given set, hazards were ranked according to each respondent's experience of recent substantial financial loss to the estate and in relation to his or her willingness to make investments aimed at risk reduction. For each hazard, the respondent assessed the risk in four classes ranging from very high to negligible risk. Six hazards were considered most problematic in all three aspects: browsing damage, falling timber prices, damage by wind, spruce bark beetle, root rot and pine weevil. A majority of the respondents claimed to take action to reduce the risk associated with at least one hazard, while 35% did not know whether they did. Excluding climate change, the need for decision support was the largest in relation to damage by wind owing to a combination of perceived high risk and a high level of ignorance in relation to whether risk-reducing measures were taken.     

Interactions in decomposition and N mineralization between tropical legume residue components

In situ produced plant residues contain a mixture of different plant components of varying quality. To assess synergistic or antagonistic interactions occurring during the decomposition and mineralization of such mixtures, individual plant parts (stems, leaves, leaf litter and roots) or the mixture of stems, leaves and leaf litter of the agroforestry species pigeonpea (Cajanus cajan) or of crop residues of peanut (Arachis hypogaea) or of the weed hairy indigo (Indigofera hirsuta) were incubated in pots for 19 weeks. Periodically, remaining plant residues were sieved out (>2 mm), weighed and N content as well as soil mineral N determined. Above- and below-ground residues of peanut decomposed fastest and showed the largest N release in agreement with their high N concentration and low-acid detergent fibre (ADF) : N ratio. Hairy indigo was hypothesized to be of lower quality than pigeonpea because of its high-polyphenol content. However, it decomposed faster than pigeonpea, largely because of the higher N and lower lignin concentration of its components. Ranking of individual plant components for N mineralization resulted in the following pattern, leaves > leaf litter > roots > stems. In mixtures of the different plant components a similar species order in decomposition was obtained, e.g. peanut > hairy indigo > pigeonpea. The amount of N released from the mixture was dominated by stem material that comprised 46–61% of the mixture. The interactions in mixtures were relatively small for peanut (generally high-quality components) as well as for pigeonpea (low proportion of high-quality components, i.e. N rich leaf material). However, a positive interaction occurred during later stages of N mineralization in the mixture of hairy indigo as it had a significant proportion of N rich components and absence of highly reactive polyphenols. Thus, for plants with low to intermediate chemical quality attributes, manipulating plant composition (e.g. by varying harvest age, affecting stem and leaf proportions) will be important to obtain significant interactions during decomposition when its components are mixed.     

Pasture production and tree growth in a young pine plantation fertilized with inorganic fertilizers and milk sewage in northwestern Spain

The use of organic waste materials such as milk sewage as an organic fertilizer could have the dual advantages of organic-waste disposal and reduced dependence on inorganic fertilizers. The effects of fertilization with (1) conventional mineral fertilization, (2) milk sewage sludge at 40 kg N ha⁻¹ target rate and (3) no fertilization on pasture production and tree growth were examined in an experiment consisting of two pasture mixtures under a one-year-old Pinus radiata plantation with a density of 2500 trees ha⁻¹. The two pasture mixtures were: (1) Dactylis glomerata L. var. saborto (25 kg ha⁻¹) + Trifolium repens L. group Ladino (4 kg ha⁻¹) + Trifolium pratense L. var. Marino (1 kg ha⁻¹); (2) Lolium perenne L. var. Tove (25 kg ha⁻¹) + Trifolium repens L. group Ladino (4 kg ha⁻¹) + Trifolium pratense L. var. Marino (1 kg ha⁻¹). The experiment began in the spring of 1995 using a randomized block design with three replicates in Castro Riberas de Lea (Lugo, Galicia, north-western Spain). Plot size was 12 × 8 m², with a 1 m buffer strip between plots. Two-year data showed that fertilization with either material had a positive effect on pasture production, with no significant difference between the two fertilization treatments. Tree growth in the milk sewage sludge plot was significantly higher than in the control plots. Inorganic fertilization increased pasture production, but affected tree growth negatively. The results show that milk sewage sludge could be used as a fertilizer in silvo-pastoral systems. This revised version was published online in June 2006 with corrections to the Cover Date.