iTRAQ protein profile analysis of soybean stems reveals new aspects critical for lodging in intercropping systems

Soybean is often intercropped with maize, sugarcane, and sorghum. Because of the shade coming from the latter, the soybean stem lodging is often a very serious problem in intercropping systems. The aim of this study is to characterize the possible mechanisms in the stem of shade-induced promotion of seedling soybean lodging in intercropping systems at the proteome level. We found that the soybean stem became slender and prone to lodging when it was planted with maize in an intercropping system. The inhibition of lignin biosynthesis and lack of photosynthate(soluble sugar) for the biosynthesis of the cell wall led to the lower internode breaking strength. A total of 317 proteins were found to be affected in the soybean stem in response to shade. Under the shade stress, the down-expression of key enzymes involving the phenylpropanoid metabolic pathway inhibited lignin biosynthesis. The up-regulation of expansin and XTHs protein expression relaxed the cell wall and promoted the elongation of internodes. Although the expression of the enzymes involving sucrose synthesis increased in the soybean stem, the lack of a carbon source prevented rapid stem growth. This metabolic deficit is the principal cause of the lower cellulose content in the stem of intercropped soybean, which leads to weakened stems and a propensity for lodging.     

Shade stress decreases stem strength of soybean through restraining lignin biosynthesis

Lodging is the most important constraint for soybean growth at seedling stage in maize-soybean relay strip intercropping system. In the field experiments, three soybean cultivars Nandou 032-4 (shade susceptible cultivar; B1), Jiuyuehuang (moderately shade tolerant cultivar; B2), and Nandou 12 (shade tolerant cultivar; B3) were used to evaluate the relationship between stem stress and lignin metabolism in the stem of soybean. Results showed that the intercropped soybean was in variable light condition throughout the day time and co-growth stage with maize. The xylem area and cross section ratio played a main role to form the stem stress. The B3 both in intercropping and monocropping expressed a high stem stress with higher xylem area, lignin content, and activity of enzymes (phenylalanine ammonia-lyase (PAL), 4-coumarate: CoA ligase (4CL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD)) than those of B1 and B2. Among the soybean cultivars and planting pattern, lignin content was positively correlated with stem stress. However, a negative correlation was found between lignin content and actual rate of lodging. In conclusion, the shade tolerant soybean cultivar had larger xylem area, higher lignin content and activities of CAD, 4CL, PAL, and POD than other soybean cultivars in intercropping. The lodging in maize-soybean intercropping can be minimized by planting shade tolerant and lodging resistant cultivar of soybean. The lignin content in stem could be a useful indicator for the evaluation of lodging resistance of soybean in intercropping and activities of enzymes were the key factors that influence the lignin biosynthesis.     

A laccase associated with lignification in loblolly pine xylem

Peroxidase has been thought to be the only enzyme that oxidizes monolignol precursors to initiate lignin formation in plants. A laccase was purified from cell walls of differentiating xylem of loblolly pine and shown to coincide in time and place with lignin formation and to oxidize monolignols to dehydrogenation products in vitro. These results suggest that laccase participates in lignin biosynthesis and therefore could be an important target for genetic engineering to modify wood properties or to improve the digestibility of forage crops.     

转录因子对木质素生物合成调控的研究进展

木质素是维管植物次生细胞壁的重要组分之一,具有重要的生物学功能。木质素分子与细胞壁中的纤维素、半纤维素等多糖分子相互交联,增加了植物细胞和组织的机械强度,其疏水性使植物细胞不易透水,利于水分及营养物质在植物体内的长距离运输。木质素与纤维素共同形成的天然物理屏障能有效阻止各种病原菌的入侵,增强了植物对各种生物及非生物胁迫的防御能力。然而木质素的存在也给人类的生产实践带来诸多负面影响,如造纸业中,由于必须使用大量化学药品去除木质素,加大了造纸成本,严重污染了环境;饲草中的高木质素含量则影响牲畜的消化吸收,降低了饲草的营养价值;过高的木质素含量也影响了人类对生物质能源的发酵利用。因此,利用基因工程改造植物木质素的可降解性意义重大。在高等植物中,木质素通过苯丙烷途径和木质素特异途径合成。在拟南芥中,NAC、MYB以及WRKY类转录因子都参与了对木质素生物合成的调控。在拟南芥中,MYB26可激活NST1/NST2的转录;WRKY12可与NST2的启动子区结合并对其表达进行负调控;SND1（NST3）和NST1主要在纤维次生壁的形成中发挥作用,两者功能有冗余;NST1和NST2在调控花药壁的次生壁的增厚中功能有冗余;VND6和VND7则主要在木质部导管的分化中起重要作用,这些NAC类转录因子通过与下游的MYB类转录因子如MYB83、MYB46及（或）MYB58、MYB63、MYB85和MYB103的结合对木质素合成基因的表达进行正调控,而MYB75对木质素生物合成进行负调控。多数MYB转录因子通过与下游木质素生物合成途径基因启动子区的AC元件（I、II和III）结合从而对其表达进行调控。研究表明,bHLH类转录因子也参与了对木质素生物合成的调控。文章综述了各类转录因子对木质素生物合成调控的最近进展,绘制了拟南芥中木质素生物合成的主要调控网络,同时也总结了其他物种（如水稻、小麦、玉米、桉树、松树和杨树等）中已发现的对木质素生物合成进行调控的转录因子。随着高通量测序技术的发展,研究者有望在更多的物种中发现参与木质素生物合成调控的关键转录因子,这些研究将对通过基因工程改造木质素的组成具有重要的借鉴意义。     

正义、反义、干涉调控肉桂酰辅酶A脱氢酶基因对烟草的影响

肉桂酰辅酶A还原酶(Cinnamoyl-CoA reductase,CCR)是在木质素合成途径中起到关键作用的酶,负责催化肉桂酰辅酶A类物质的还原反应生成相应的醛类化合物.分离得到毛白杨CCR基因的cDNA.将CCR基因以正义、反义和干涉的形式,通过农杆菌介导的方法转入烟草中,Southem杂交以确定转基因成功.在获得的转基因植株中,所有下调基因的植株都表现出可溶性酚酸含量下降,木质素含量也有明显的下降趋势,纤维素含量则有上升,可能是对木质素含量下降的一种代偿性增长.植物细胞壁组分的含量变化表明CCR基因可能是一个控制木质素代谢途径的一个很好的调控点.     

不同年限设施菜地番茄细胞壁果胶Cd累积的研究

为探讨不同种植年限和设施菜地番茄细胞壁果胶Cd累积的关系,以种植年限为2年和14年设施菜地番茄为研究对象,测定了不同种植年限下番茄茎、叶细胞壁果胶含量,并对细胞壁果胶含量、果胶甲酯酶(PME)活性与种植年限及Cd累积量之间的相关性进行了分析。结果表明,随着种植年限的增加,番茄植株各器官Cd累积量均有所增加;种植14年设施菜地番茄茎中Cd累积量比种植2年的增加了24.55%,叶片中Cd累积量增加了45.96%;种植14年设施菜地番茄茎细胞壁Cd累积量增加了61.86%,叶片细胞壁Cd累积量增加了57.46%;不同种植年限设施菜地番茄茎和叶细胞壁果胶Cd含量也存在极显著差异(P0.05)。另外,随着种植年限的增加,番茄茎、叶细胞壁果胶含量和PME活性随之升高:与种植2年相比,种植14年设施菜地番茄茎中果胶含量提高了104.51%,叶片果胶含量提高了127.45%;种植14年设施菜地番茄茎中PME活性提高了10.56%,叶片PME活性提高了9.39%。果胶含量及PME活性和细胞壁Cd累积量呈正相关。研究发现不同年限设施菜地土壤全Cd和有效Cd含量有所不同,影响着番茄不同器官果胶含量及PME活性,说明细胞壁果胶在缓解番茄Cd毒害中起着一定的作用。     

过表达GhMYB4基因对棉花茎秆木质素合成的影响

旨在探究过表达陆地棉(Gossypiumhirsutum)GhMYB4基因对棉花茎秆木质素合成的影响。利用Wiesner组织化学染色的方法对棉花茎秆进行染色,并用Klason法对棉花茎秆的局部位置进行木质素总量测定。对距离下胚轴0～1 cm、2～7cm、10～12 cm处不同茎秆部位的次生壁木质素进行化学组织染色,定量分析染色组织结构发现,大部分的转基因株系相比对照受体株系在初生木质部和初生韧皮部的染色范围、细胞层数和染色程度上均有一定的显著性增加。通过对29个过表达转基因株系和对照受体株系在棉花茎秆距离下胚轴2～7cm处木质素总量的定量分析发现,有23个过表达转基因株系相比于对照受体株系的木质素总量有显著性的增加和1个株系出现显著性降低的现象。这些结果表明GhMYB4基因参与调控棉花茎秆的木质素生物合成。     

遮阴胁迫对丹参生长及光合特性的影响

[目的]研究不同遮阴胁迫对丹参生长及光合特性影响。[方法]通过在不同遮阴处理条件下对丹参植株高度、小叶片面积、叶片数等主要农艺性状的调查以及丹参主要生育期光合速率的测定,分析遮阴胁迫对丹参生长及光合特性的影响。[结果]在不同遮阴条件下,随着光照强度的减弱,对丹参生长的影响主要表现在茎变细,植株的高度、小叶片面积、叶片数先增加后减少。在全光照条件下,过强的光照不利于丹参的生长。7、8月份在海棠地2.0 m×3.0 m的遮阴处理下即郁闭度为32.7%时叶面积最大,叶片开展好,且药用器官根增粗增长,说明适度遮阴有利于丹参植株的营养生长。[结论]处于全光照和郁闭度32.7%之间的遮阴适于丹参植株的生长。     

植物扩展蛋白基因的研究进展

扩展蛋白基因(expansin genes)是一个相对保守的基因家族,由α、β、γ和δ4个亚家族组成。作为植物细胞壁的重要组成部分,扩展蛋白以酶催化的作用方式,使细胞壁组分间疏松,细胞伸展,细胞的柔韧性增强,以此缓解各种环境因子对细胞的压力。目前研究发现:扩展蛋白参与了植物生长发育过程中的许多环节,如种子萌发、根毛的起始和延长、叶子生长发育、叶柄脱落、花粉管延长、果实成熟等。同时,扩展蛋白还参与了抗旱、抗病、耐高温等诸多抗逆性反应。扩展蛋白基因多属于诱导表达,易受激素、温度、干旱、病原菌、机械损伤等各种内外界环境因素的影响。