Activities of chitinolytic enzymes during primary and secondary colonization of wood by basidiomycetous fungi

The nitrogen (N) content of wood is usually suboptimal for fungal colonization. During decomposition of wood, an increasing fraction of the N becomes incorporated into fungal mycelium. Between 5 and 50% of the N in wood-degrading mycelium may be incorporated into chitin. Chitinolytic enzymes render this N available for re-utilization. Here, the activities of chitinolytic enzymes produced by wood-rotting fungi during degradation of spruce (Picea abies) wood were quantified in situ using fluorogenic 4-methylumbelliferyl substrates. A new method was developed that enables spatial quantification of enzyme activities on solid surfaces. All of the three tested fungi produced endochitinases, chitobiosidases and N-acetylhexosaminidases during colonization of wood. N-acetylhexosaminidase activity, and in some cases also chitobiosidase and endochitinase activities, were higher during secondary overgrowth of another fungus than during primary colonization of noncolonized wood. The results suggest that wood-degrading fungi degrade their own cell walls as well as the hyphae of earlier colonizers. Recycling of cell wall material within single mycelia and between fungal individuals during succession may lead to retention of N within woody debris.     

Towards a dynamic photosynthesis model to guide yield improvement in C4 crops

The most productive C4 food and biofuel crops, such as Saccharum officinarum (sugarcane), Sorghum bicolor (sorghum) and Zea mays (maize), all use NADP-ME-type C4 photosynthesis. Despite high productivities, these crops fall well short of the theoretical maximum solar conversion efficiency of 6%. Understanding the basis of these inefficiencies is key for bioengineering and breeding strategies to increase the sustainable productivity of these major C4 crops. Photosynthesis is studied predominantly at steady state in saturating light. In field stands of these crops light is continually changing, and often with rapid fluctuations. Although light may change in a second, the adjustment of photosynthesis may take many minutes, leading to inefficiencies. We measured the rates of CO₂ uptake and stomatal conductance of maize, sorghum and sugarcane under fluctuating light regimes. The gas exchange results were combined with a new dynamic photosynthesis model to infer the limiting factors under non-steady-state conditions. The dynamic photosynthesis model was developed from an existing C4 metabolic model for maize and extended to include: (i) post-translational regulation of key photosynthetic enzymes and their temperature responses; (ii) dynamic stomatal conductance; and (iii) leaf energy balance. Testing the model outputs against measured rates of leaf CO₂ uptake and stomatal conductance in the three C4 crops indicated that Rubisco activase, the pyruvate phosphate dikinase regulatory protein and stomatal conductance are the major limitations to the efficiency of NADP-ME-type C4 photosynthesis during dark-to-high light transitions. We propose that the level of influence of these limiting factors make them targets for bioengineering the improved photosynthetic efficiency of these key crops.     

新疆荒漠植物耳叶补血草的爆发式开花式样与传粉模式

荒漠环境中干热、大风、浮尘、温度骤变等突发性天气状况,对植物的有性生殖具不利影响.耳叶补血草(Limonium otolepis)是新疆盐生荒漠的主要建群种,对其适应荒漠恶劣环境的生殖策略仍缺乏研究.为了探讨荒漠植物适应环境的开花式样和传粉模式,作者通过野外定点观测和室内电镜扫描等方法对耳叶补血草的开花进程、花部特征、传粉媒介、结实特性等进行了研究.耳叶补血草的花期在6月上旬至7月中旬,花期较长且具明显不同步的单株开花状态.在一天中,单花表现出极为集中的开花、散粉过程:一般早上8:00开始开放,9:00达开放高峰期,11:00以后基本不再有单花开放;开放后的单花持续6-8 h后闭合.每单花平均有花粉752粒,花粉表面有网状纹饰,有花蜜.传粉者主要为蜜蜂科和食蚜蝇科昆虫,访花高峰期在9:00-14:00.人工授粉实验表明耳叶补血草以异交为主,自动自花授粉率低.自然状态下结实率为36％,在花序中基部第一位上单花的结实率(45.7％)明显高于第二位上单花(3.3％).耳叶补血草爆发式开花、在短时间内快速完成传粉过程,可能是躲避荒漠环境突发性天气条件的一种策略;而居群分散的开花时间及较长的花期在应对不稳定生境,分摊生殖风险方面具重要意义.     

盐碱胁迫对两种高粱种子萌发及幼苗生长的影响

以纤维高粱和普通高粱种子为材料,将NaCl、Na2SO4和NaHCO3按不同比例混合,模拟15种盐度和碱度各不相同的复杂盐碱条件,采用培养皿纸上发芽法研究了混合盐碱胁迫对两种高粱种子萌发及幼苗生长的影响,以探讨两种高粱种子和幼苗的耐盐碱能力及在盐碱胁迫下的变化规律。结果表明,低盐低碱环境促进了两种高粱种子的萌发;随着盐碱胁迫的增强,高粱种子的萌发率、萌发指数与对照相比均显著下降,而纤维高粱的累积萌发率下降幅度较普通高粱小,表明纤维高粱对盐渍环境的适应性略高于普通高粱。在高粱幼苗生长过程中,随着盐碱胁迫的增强,两种高粱的幼根和幼芽长均显著下降,且与对照均具有显著性差异。盐度对两种高粱的萌发率、萌发指数、幼苗长均有极显著影响,盐度与pH相互作用对两种高粱的幼根长有极显著影响。     

A combined biochemical screen and TILLING approach identifies mutations in Sorghum bicolor L. Moench resulting in acyanogenic forage production

Cyanogenic glucosides are present in several crop plants and can pose a significant problem for human and animal consumption, because of their ability to release toxic hydrogen cyanide. Sorghum bicolor L. contains the cyanogenic glucoside dhurrin. A qualitative biochemical screen of the M2 population derived from EMS treatment of sorghum seeds, followed by the reverse genetic technique of Targeted Induced Local Lesions in Genomes (TILLING), was employed to identify mutants with altered hydrogen cyanide potential (HCNp). Characterization of these plants identified mutations affecting the function or expression of dhurrin biosynthesis enzymes, and the ability of plants to catabolise dhurrin. The main focus in this study is on acyanogenic or low cyanide releasing lines that contain mutations in CYP79A1, the cytochrome P450 enzyme catalysing the first committed step in dhurrin synthesis. Molecular modelling supports the measured effects on CYP79A1 activity in the mutant lines. Plants harbouring a P414L mutation in CYP79A1 are acyanogenic when homozygous for this mutation and are phenotypically normal, except for slightly slower growth at early seedling stage. Detailed biochemical analyses demonstrate that the enzyme is present in wild-type amounts but is catalytically inactive. Additional mutants capable of producing dhurrin at normal levels in young seedlings but with negligible leaf dhurrin levels in mature plants were also identified. No mutations were detected in the coding sequence of dhurrin biosynthetic genes in this second group of mutants, which are as tall or taller, and leafier than nonmutated lines. These sorghum mutants with reduced or negligible dhurrin content may be ideally suited for forage production.     

Morphological and molecular characterization of sweet,grain and forage sorghum (Sorghum bicolor L.) genotypes grown under temperate climatic conditions

Abstract

In the present study, we used 12 genotypes of sorghum originated from different countries (five sweet, four grain and three forage). These different genotypes and types of sorghum were evaluated for the agro-morphological traits that are associated with the estimated sugar and bioethanol yield to estimate their phenotypic diversity. Analysis of variance showed significant differences between different types of sorghum for all the evaluated traits. Sweet sorghum genotypes, however, showed better performance with respect to all studied traits than the other genotypes. A positive significant correlation was observed between plant height, leaf number, leaf area, biomass yield, cane and bagasse yields, and the predicted bioethanol yield. Both, cluster and principal component analysis were performed to group the genotypes according to their agro-morphological and molecular similarity coefficients. For analytical approaches, the Iranian grain and forage genotypes clustered separately from the other genotypes. The clustering patterns obtained from the molecular dominant markers had higher discriminatory power than using morphological characters to separate sweet genotypes from the forage and grain sorghum ones. The results clearly indicated that sweet sorghum can be grown in Germany and maintains its superiority in biomass production and sugar yield over grain and forage sorghum types.     

高梁CRY2基因的克隆及其表达分析

从高粱(Sorghum bicolor L. var.R1ll)幼苗中提取总RNA,利用RT-PCR和cDNA的3'末端的快速扩增方法(3'RACE),第一次克隆了高粱隐花色素2基因(CRY2)的cDNA序列.该序列包括了一个完整的开放阅读框,编码大小为690个氨基酸残基的蛋白质,与水稻、番茄和拟南芥CRY2蛋白质的同源性分别为8 7%、5 7%和45.5%.高粱CRY2基因组DNA含有3个内含子和4个外显子.RT-PCR检测结果表明,高粱CRY2基因在根、茎和叶中都有转录.Western blotting结果显示CRY2蛋白在根、茎和叶中表达,并在黑暗中积累,蓝光下降解.高粱CRY2可能在蓝光诱导的幼苗去黄化反应中起作用.