设施栽培中桃树14C同化物分配特性的研究

 利用放射性同位素¹⁴C - 示踪方法研究了设施桃树果实不同发育时期¹⁴ C同化物的运转分配特性。结果表明: 在果实膨大期和果实成熟期, 分配到果实中的¹⁴C同化物最多, 随着果实生长, 分配到果实中的¹⁴C同化物比例增加。叶片的自留量小于果实获得的¹⁴C同化物量, 且随着果实的生长, 叶片的自留量逐渐减少。在各器官中果实的放射比活性最强, 其次为叶片与根系。说明果实膨大期和果实成熟期是设施桃树对碳素同化物需求和竞争最大的时期。     

荔枝果皮对外源钙和蔗糖吸收及向细胞壁沉着的研究

利用放射性同位素示踪技术研究荔枝果皮对外源钙和蔗糖的吸收及细胞壁构建规律。结果表明,进入果皮的蔗糖主要被用以构建细胞壁,但随果实发育,蔗糖用于构建细胞壁比例减少,更多分布于果皮组织的可溶性成分中;果梗向果实运输钙的效率远远低于蔗糖运输效率;NAA处理果实短期内可明显促进蔗糖向果实运输,但不能促进果梗的钙进入果实,说明蔗糖和钙向果实运输有不同调控机制;施于果实表面的钙虽可被吸收并成为细胞壁的结构钙,但比例不到千分之一;硝酸(根)离子和NAA可一定程度促进外源钙向细胞壁沉着;抗裂的怀枝果皮细胞壁的钙含量比易裂的糯米糍高,其细胞壁结合外源钙的能力也强于后者,说明前者细胞壁中果胶半乳糖醛酸残基含量高于后者,也是怀枝具有较强抗裂性的物质基础之一;2品种14C-蔗糖向果皮细胞壁沉着均在果实发育初期最活跃,怀枝细胞壁构建也只是在果实发育初期比糯米糍更活跃,这就意味着抗裂性形成的关键时期是在果皮发育的初期。     

伪狂犬病毒UL14基因缺失突变株的构建及其生物学特性研究

UL14在α-疱疹病毒中相当保守,但其在伪狂犬病毒中的功能尚不清楚。本研究以伪狂犬病毒鄂A株（PRV-Ea）为亲本株,通过同源重组,将编码EGFP的真核表达盒插入到伪狂犬病毒基因组的UL14基因中,PCR检测和荧光观察证实获得了能稳定表达EGFP的UL14插入失活突变株PRV-UL14D。将PRV-UL14D感染IBRS-2细胞,发现该突变株较PRV-Ea野毒株增殖缓慢,而且PRV-UL14D突变株感染细胞后形成的蚀斑也明显小于PRV-Ea野毒株,但在稳定表达UL14的IBRS-2细胞系中,PRV-UL14D与PRV-Ea野毒株的增殖速度、形成的蚀斑大小均无明显差异。进一步将PRV-UL14D感染BALB/c小鼠,发现与相同剂量的PRV-Ea野毒株感染相比,小鼠平均死亡时间明显延缓。上述研究结果表明,UL14并非伪狂犬病毒增殖必需的,但其缺失可延缓病毒增殖,推测UL14可能与病毒粒子在细胞间的扩散有关。     

Cyclopedic protein expression analysis of cultured canine mammary gland adenocarcinoma cells from six tumours

We characterised cultured canine mammary gland adenocarcinoma cells by exhaustive step protein expression analysis to identify factors associated with tumour progression or metastasis of canine mammary gland tumour. Cultured adenocarcinoma cells derived from a total of 3 primary and 3 metastatic lesions from 3 dogs (CHMp/m, CIPp/m and CNMp/m, where CHM, CIP, and CNM indicate the 3 animals) were used in this study. The expression of 24 proteins reported to be related to tumourigenesis or malignancy of human breast cancers were examined by Western blot analysis using 24 antibodies. The expression of sialyl Lewis X [sLe(x)] was only observed in CHMm cells, which were derived from pleural effusion. This expression was further confirmed by immunohistochemistry. The levels of some factors, such as 14-3-3sigma, cyclinD1 and Rb, differed among cells or between the primary and metastatic cells in the pair. Though the difference in their expression was not consistent within the cells from primary and metastatic origin, this characterisation should provide useful information for further molecular analysis of these cultured cells. Since some of the factors, such as sLe(x), 14-3-3sigma, cyclinD1 and Rb, showed different levels of expression in the pair, these cultured cells might be meaningful tools for clarification of distant metastasis in canine mammary gland tumours.     

蛇床子素对小麦赤霉病菌葡萄糖、钙吸收和三磷酸腺苷酶活性的抑制

蛇床子素可抑制小麦赤霉病菌菌丝生长干重;含有14C标记的葡萄糖培养基中培养小麦赤霉病菌,以100 μg/mL蛇床子素处理,其菌体内14C放射强度比对照降低43％,表明蛇床子素能抑制小麦赤霉病菌对葡萄糖的吸收;以100 μg/mL蛇床子素处理小麦赤霉病菌12 h,菌体内总钙含量仅为对照的60%,表明蛇床子素能抑制菌体对钙的吸收;蛇床子素处理对β-1,6-葡聚糖酶活性没有显著影响;离体条件下,蛇床子素能抑制小麦赤霉病菌三磷酸腺苷(ATP)酶活性。     

Polymorphism of 14α-demethylase Gene (CYP51) in the Cereal Eyespot Fungi Tapesia acuformis and Tapesia yallundae

Cereal eyespot fungi Tapesia acuformis and Tapesia yallundae are closely related species which show different behaviours upon treatment with sterol 14-demethylase inhibitors (DMIs). T. acuformis is naturally resistant to DMIs belonging to the triazole family and susceptible to the imidazole ones, whilst T. yallundae is sensitive to both inhibitors. Cloning of the target enzyme gene, CYP51, from the two species revealed an important polymorphism between them. Further sequencing of CYP51 from sixteen T. acuformis and eleven T. yallundae strains with different phenotypes with regards to resistance to DMIs confirmed that at least eleven variations are species related. Among them, a conserved phenylalanine residue at position 180, found both in T. yallundae and in all known CYP51 proteins from filamentous fungi and yeast, was replaced in T. acuformis by a leucine. Therefore, a leucine at 180 could be possibly involved in natural resistance of T. acuformis to triazoles. Other mutations were observed in some resistant strains, sometimes simultaneously, but in contrast to what was reported for other filamentous fungi, where a mutation at the 136 position of the CYP51 gene product seemed to correlate with resistance to DMIs, we did not find a clear relationship between a given mutation and a particular phenotype. This result suggests that resistance to DMIs could have a polygenic nature in Tapesia. We took advantage of species-related variations to develop a PCR-based assay allowing rapid and easy discrimination between field strains of the two species.     

大白菜种株碳同化分配及其对种荚发育的作用

用　14 C示踪方法研究大白菜种株不同生育时期同化产物分配及其对种荚形成的作用 ,结果表明 :叶片是主要同化器官 ;生育前期同化产物主要向茎枝和根系分配 ;后期主要向角果输送 ;种株开花前同化产物可通过再次分配进入角果 ,开花后同化产物对种子形成贡献更大。种株不同生育时期同化产物对角果皮和籽粒发育的作用存在差异     

IAA、GA和ABA对葡萄果实~(14)C蔗糖输入与代谢的调控

将　14 C蔗糖与激素IAA、GA和ABA同时通过果柄引入离体葡萄果实 ,研究了蔗糖积累代谢与激素的关系。结果表明 :①GA在幼果膨大期 ,IAA在果实始熟期以前 ,ABA从缓慢生长期到果实成熟 ,对蔗糖的吸收有明显促进效果。②GA与IAA在果实发育前期 (第Ⅰ、Ⅱ期 )可促进　14 C蔗糖转化为非醇溶性结构物 ;ABA则在成熟期表现出类似的作用。③IAA、GA与ABA在幼果膨大期可促进糖分转化为有机酸 ,但在缓慢生长期都抑制这一过程 ,抑制效果以ABA最显著 ,IAA其次 ,GA最小。成熟期糖分占醇溶物的 85 %～ 91% ,不同激素处理的差异不明显。ABA可促进各个时期果实中氨基酸的合成。④IAA可加快蔗糖分解为还原糖 ;GA增加果糖积累 ;ABA处理的果实蔗糖含量最高 ,推测ABA直接刺激了果肉维管束的蔗糖卸载。     

Characterisation of energy-dependent efflux of imazalil and fenarimol in isolates of Penicillium italicum with a low,medium and high degree of resistance to DMI-fungicides

Differential accumulation of [¹⁴C]imazalil and [¹⁴C]fenarimol by germlings of wild-type and DMI-resistant isolates ofPenicillium italicum was studied at various pH values. At pH 7 and 8 the low-resistant isolate E_300–3 accumulated 22% and 35%, respectively, less imazalil than the wild-type isolate W₅. Imazalil accumulation at pH 5 and 6 was similar. Isolate E_300–3 also accumulated less fenarimol as compared with the wild-type isolate. This difference was much more obvious than for imazalil and was observed at all pH values tested. Differences in accumulation of both imazalil and fenarimol between low (E_300–3), medium (H₁₇) and high resistant (I₃₃) isolates were not observed. These results suggest that decreased accumulation of DMIs is responsible for a low level of resistance only and that additional mechanisms of resistance might operate in isolates with a medium and high degree of resistance. With all isolates fenarimol accumulation was energy-dependent. This was not obvious for imazalil.The wild-type and DMI-resistant isolates had a similar plasma membrane potential as determined with the probe [¹⁴C]tetraphenylphosphonium bromide ([¹⁴C]TPP⁺). Various test compounds, among which ATPase inhibitors, ionophoric antibiotics and calmodulin antagonists, affected the accumulation of [¹⁴C]TPP⁺, [¹⁴C]imazalil and [¹⁴C]fenarimol. No obvious correlation between the effects of the test compounds on accumulation levels of the fungicides and [¹⁴C]TPP⁺ could be observed. These results indicate that the plasma membrane potential does not mediate the efflux of DMI fungicides byP. italicum.     

梭梭14-3-3蛋白基因HaFT-9的克隆及特性分析

【目的】分析梭梭14-3-3蛋白基因HaFT-9在不同胁迫下的表达模式,解析HaFT-9蛋白结合特性,为研究梭梭抗逆分子机制提供理论基础。【方法】以梭梭cDNA为模板克隆14-3-3蛋白基因HaFT-9,利用qRT-PCR技术对该基因在不同胁迫下的表达模式进行分析,并通过酵母双杂交实验验证HaFT-9蛋白结合特性。【结果】克隆得到ORF长度为789 bp的梭梭14-3-3蛋白基因,命名为HaFT-9。梭梭幼苗在20% PEG6000、200 mmol/L NaCl和100 μmol/L ABA处理下,HaFT-9的表达量均在胁迫6 h时达到峰值,在20 μmol/L IAA处理下,HaFT-9的表达量在胁迫1 h时达到峰值。梭梭幼苗经40、55℃和55、65℃模拟地表高温胁迫,高温胁迫适应性试验中的常温复原阶段HaFT-9均上调表达。酵母双杂交实验表明HaFT-9可与自身形成同源二聚体。【结论】克隆得到HaFT-9基因(登录号为API85525.1),该基因可响应干旱胁迫、盐胁迫、ABA和IAA处理,并与高温胁迫适应性相关,HaFT-9蛋白能与自身互作形成同源二聚体。