Molecular Cloning of a Cellulose Synthase Gene PtoCesA1 from Populus tomentosa and Its Genetic Transformation in Tobacco

A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P tomentosa. Four anti-expression vectors with different fragments of PtoCesAl, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, ＂dwarf＂ phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems.     

转基因741杨对舞毒蛾不同龄期幼虫的抗性研究

用7种不同系号的转抗741叶片为材料，对舞毒蛾幼虫进行逐龄分别饲养，系统测定各系号抗虫杨叶片对舞毒蛾不同龄期幼虫的死亡率，体重增长速率，发育速率的影响。结果表明：7种抗虫杨叶片对1龄幼虫均表现出较高的毒杀作用，其中处理3d的死亡率均达到35％，处理5d死亡率达80％。对2，3龄幼虫除具有一定的毒杀作用外，主要表现为对其生长发育的抑制作用，影响其正常的取食，降低体重增长速率和减慢发育速率。     

转多基因库安托杨非靶目标材性性状分析

In this study, the transgenic poplar (Populus × euramericana‘Guariento’) harboring five exogenous genes (SacB, Vgb, BtCry3A, OC-I and JERF36^#) and the control plants were selected, comparing the wood property variation of 5-year-old trees, and trying to understand whether the transgenic of exogenous genes produce unintended effects. The results showed that the transgenic of exogenous genes caused a wide range of variation in non-target wood properties, the variations were found not only between transgenic and wild-type lines, but also among transgenic lines. Compared with the control, the transgenic lines, the contents of holocellulose and cellulose increased, the total lignin and acid soluble lignin decreased, the fiber length, the fiber width, the microfibril angle and the acid insoluble lignin varied uncertainly. It is concluded that the introduced exogenous gene could generate unintended effects on corresponding non-target target traits.     

余甘果抗衰老作用的研究

选用果蝇作实验动物，通过余甘原汁、余甘粉和余甘营养液对果蝇寿命影响的研究表明，在一定的浓度范围内，余甘子的３种产品都能延长果蝇寿命，尤其是２５％浓度余甘营养液能使果蝇平均寿命比对照延长１倍以上，从而进一步验证了余甘果的抗衰老作用。     

转双抗虫基因741杨嫩枝扦插苗抗寒性的研究

为研究转双抗虫基因741杨不同时期扦插苗的抗寒性,以5月、6月、7月、8月嫩枝扦插苗为试验材料,进行低温处理,测定生理反应和恢复生长能力。结果表明,7月、8月嫩枝扦插苗可溶性蛋白质含量低,-15℃以下低温相对电导率明显增高,枝条皮部逐渐变黑,芽失去萌发能力;而5月、6月嫩枝扦插苗对寒冷适应性较强,-15℃以下低温仍具一定恢复生长能力。     

掺钢渣的石灰粉煤灰稳定碎石抗压强度的研究

以马鞍山钢铁集团钢渣公司的钢渣为研究对象，分析研究了在半刚性基层材料石灰粉煤灰稳定碎石中掺入细钢渣，用细钢渣部分取代粉煤灰后，混合料无侧限抗压强度的变化情况。试验结果表明，随着细钢渣掺量的增加，石灰粉煤灰稳定碎石混合料的无侧限抗压强度呈先增大后减小的趋势。这说明当细钢渣掺量适当时（钢渣：粉煤灰=1：2），能明显提高石灰粉煤灰稳定碎石混合料的抗压强度，其抗压强度可提高30％，所以从其增强效果来看是比较显著的。     

基于宿主诱导的基因沉默技术创制抗拟轮枝镰孢玉米自交系

【目的】拟轮枝镰孢（Fusarium verticillioide）是一种主要的病原真菌,侵染玉米可以导致穗粒腐病、茎腐病、苗期根腐病及引起种子腐烂。由拟轮枝镰孢引起的病害不仅影响玉米的产量和品质,而且其病原菌代谢过程中产生的伏马菌素等多种真菌毒素严重威胁了人畜安全。通过宿主诱导的基因沉默（host-induced gene silencing,HIGS）技术创制抗拟轮枝镰孢的玉米种质,为玉米抗病育种提供新的优异抗源。【方法】通过同源克隆方法克隆可能与拟轮枝镰孢生长发育相关的关键基因,并通过体外转录获得相应的dsRNA片段;将不同基因的dsRNA与拟轮枝镰孢的分生孢子悬浮液预混后,用于后续的体外RNA沉默试验;对感病玉米自交系西502的种子进行消毒与接种,在培养皿中28℃避光培养48 h,调查种子的发病程度;在混有dsRNA的孢子悬浮液中加入葡萄糖,25℃培养24 h后,在显微镜下观察孢子萌发与菌丝生长情况;将三叶期的西502幼苗转移至预混dsRNA的孢子悬浮液中进行培养,7 d后观察苗期根腐病发病状况;通过种子鉴定与苗期鉴定体系,逐步筛选具有显著抑制效果的沉默靶标基因;合成筛选出的重点靶标基因片段,构建沉默载体并转化感病玉米自交系西502;对转基因株系的种子接种鉴定,验证转化玉米株系的抗性;提取接种后转基因种子的总RNA,对拟轮枝镰孢的靶标基因进行荧光定量分析,确定HIGS株系的沉默效果。【结果】从拟轮枝镰孢中克隆出18个与其生长发育相关的候选基因;通过种子接种鉴定,发现11个候选基因被沉默后,种子的发病等级极显著降低;进一步筛选出6个沉默后影响拟轮枝镰孢的孢子萌发和菌丝生长的候选靶标基因deo、Ras2、Dpdc、Hsp90、Frp1和Atg15;通过苗期接种鉴定,最后筛选出3个在体外具有显著抑制效果的沉默靶标基因deo、Atg15和Frp1;进而将3个靶标基因的特异区段人工融合成一段序列并构建沉默载体,获得转基因植株;鉴定发现转基因植株的T₂代种子对拟轮枝镰孢的抗性显著增强,且3个靶标基因的表达量均显著下降。【结论】拟轮枝镰孢基因deo、Atg15、Frp1与其生长发育密切相关,且沉默后能够显著提升玉米对拟轮枝镰孢的抗性。     

狐貉粪对黑水虻生长发育的影响

为了研究狐貉粪对黑水虻幼虫生长、预蛹及蛹的发育、成虫历期、饲料系数的影响,在温度20~25℃、湿度70%~80%的环境下,以狐貉粪为主料,添加不同比例的基础料（麦麸、豆粕与玉米面）进行饲喂。结果表明：30%发酵混合料+70%基础料处理下,老熟幼虫、预蛹、蛹的质量分别为156.0、108.8、60.2 mg,显著高于对照组,预蛹历期、蛹历期分别为7、6 d,相较于对照组显著缩短;30%发酵混合料+70%基础料的黑水虻成活率、化蛹率、羽化率都较高,分别为93%、88%、91%;50%发酵混合料+50%基础料处理组除幼虫体重显著低于30%发酵混合料+70%基础料处理组外,其他指标差异均不显著。综合分析认为,30%~50%发酵混合料对黑水虻生长发育最为适宜。     

种植密度对转Bt棉纤维杀虫蛋白表达量及氮代谢的影响

为了探讨种植密度对转基因抗虫棉纤维中杀虫(Bt)蛋白表达量的影响及其氮代谢机制,以Bt棉常规种泗抗1号和Bt棉杂交种泗抗3号为试验材料,比较1.5×10⁴、3.0×10⁴、4.5×10⁴、6.0×10⁴、7.5×10⁴ 株·hm^-2五个种植密度下转Bt棉杀虫蛋白表达量及其氮代谢的差异。结果表明,纤维中Bt蛋白表达量随种植密度增大呈上升趋势。相关分析表明,单株铃数、单铃体积、单铃干重与Bt蛋白表达量均呈显著负相关。纤维中氮代谢生理相关指标中,可溶性蛋白含量及谷氨酸丙酮酸转氨酶(GPT)和谷氨酸草酰乙酸转氨酶(GOT)活性随种植密度的增大而升高,但游离氨基酸含量、蛋白酶和肽酶活性随密度增大而下降。纤维中Bt蛋白表达量与可溶性蛋白含量、GPT和GOT活性呈显著正相关,而与游离氨基酸含量、蛋白酶和肽酶活性呈显著负相关。可见,提高种植密度降低了棉铃生长,但提高了纤维中Bt蛋白的表达量,且Bt蛋白表达量的提高与植株氮代谢生理密切相关。因此,在生产中通过协调种植密度既可以保持棉铃适度发育,又有利于提高棉纤维中Bt蛋白的表达量,为提高Bt棉产量器官抗虫性提供栽培基础。     

Transgenic woody plants for biofuel

Transgenic trees as a new source for biofuel have brought a great interest in tree biotechnology. Genetically modifying forest trees for ethanol production have advantages in technical challenges, costs, environmental concerns, and financial problems over some of crops. Genetic engineering of forest trees can be used to reduce the level of lignin, to produce the fast-growing trees, to develop trees with higher cellulose, and to allow the trees to be grown more widely. Trees can establish themselves in the field with less care of farmers, compared to most of crops. Transgenic crops as a new source for biofuel have been recently reviewed in several reviews. Here, we overview transgenic woody plants as a new source for biofuel including genetically modified woody plants and environment; main focus of woody plants genetic modifications; solar to chemical energy transfer; cellulose biosynthesis; lignin biosynthesis; and cellulosic ethanol as biofuel.