木质部细胞分化的研究进展

本文主要从研究木质部细胞分化常用的实验系统,木质部分的诱导,木质部细胞的编程性死亡以及次生壁的构建4个方面阐述了木质细胞分的研究进展,并对目前研究的热点也是难点问题进行了展望,希望引起同行的兴趣。     

木质部细胞分化和脱分化的机理

木质部细胞的分化过程包括了密切不可分的细胞程序死亡和次生壁构建两个过程。现在的研究主要是将两个过程分开来研究,各自在细胞生物学和分子生物学上取得了不少进展,有关次生细胞壁方面的研究时间长,成果也较大。有关木质部细胞脱分化的研究相对较少,但也已取得了可喜的进展。     

木质部细胞分化的程序

本文主要对近十几年来有关木质部细胞分化研究中使用的实验系统及用这些系统所取得的重要进展作了评述.并以作者实验室的研究成果为基础,结合国内外研究进展,提出木质部细胞分化程序由参与细胞编程死亡(PCD)和次生壁构建的全部基因综合编制而成.以PCD过程各阶段的划分标准来看,木质部细胞分化中从IAA诱导形成层细胞平周分裂到细胞扩大前为PCD的起始阶段,其间包括死亡信号的发生、接受和传导,以及启始caspase(半胱氨酰基天门冬氨酸蛋白酶)类似物(例如caspase-8类似物)的活化;木质部母细胞的径向扩大为PCD的效应阶段,而效应caspase类似物(例如caspase-3类似物)活化DNase、DNA的片段化及次生细胞壁的构建和各种细胞器的解体则为PCD的清除降解阶段.至今还无法将DNase活化及其引起的DNA断裂过程与次生细胞壁构建过程分开.     

杜仲次生木质部细胞分化和脱分化过程中ATPase的超微细胞化学定位

采用磷酸铅沉淀技术,对杜仲（Ｅｕｃｏｍｍｉａ ｕｌｍｏｉｄｅｓ Ｏｌｉｖ．）次生木质部细胞分和脱化过程进行了ＡＴＰａｓｅ的超微细胞化学定位。随着分化过程中细胞程序性死亡（ｐｒｏｇｒａｍｍｅｄ ｃｅｌｌ ｄｅａｔｈ,ＰＣＤ）程度的加深,ＡＴＰａｓｅ在细胞核上的分布由少变多,而在各种细胞器上的分布由有到无,并且随着细胞质的解体,ＡＴＰａｓｅ在细胞壁内侧和纹孔处的分布也由少到多,说明它们的变化是由核基因     

杜仲次生木质部分化过程中的细胞编程死亡

通过电子显微镜观察、DNA断裂检测及类似半胱氨酸蛋白酶（caspase-like proteases,CLPs)降解检测等技术,对杜仲（Eucommia ulmoides Oliv.）次生木质部分化过程的细胞编程死亡进行了研究。分化中的次生木质部细胞总DNA凝胶电泳检测到DNA ladder,并通过TUNEL检测进一步确定了DNA被降解。Western blot结果表明：caspase-8和caspase-3状蛋白酶（caspase-8-和caspase-3-like proteases,CLPs)及多聚ADP-核糖聚合酶（poly(ADP-ribose) polymerase,PARP)在次生木质部分化过程中被降解。这些研究结果表明,杜仲次生木质部的细胞分化是一个典型的编程性死亡（Programmed cell death,PCD)过程,CLPs可能参与了此过程。     

杜仲次生木质部分化过程中的细胞编程死亡

通过电子显微镜观察、DNA断裂检测及类似半胱氨酸蛋白酶(caspase-like proteases,CLPs)降解检测等技术,对杜仲(Eucommia ulmoides Oliv.)次生木质部分化过程的细胞编程死亡进行了研究.分化中的次生木质部细胞总DNA凝胶电泳检测到DNA ladder,并通过TUNEL检测进一步确定了DNA被降解.Western blot结果表明;caspase-8和caspase-3状蛋白酶(caspase-8-和caspase-3-like proteases,CLPs)及多聚ADP-核糖聚合酶(poly(ADP-ribose)polymerase,PARP)在次生木质部分化过程中被降解.这些研究结果表明,杜仲次生木质部的细胞分化是一个典型的编程性死亡(Programmed cell death,PCD)过程,CLPs可能参与了此过程.     

杜仲次生木质部分化过程中木质素与半纤维素组分在细胞壁中分布的动态变化

利用紫外光显微镜、透射电子显微镜结合免疫胶体金标记,研究了杜仲（Eucommia ulmoides Oliv.)次生木质部分化过程中木质素与半纤维素组分（木葡聚糖和木聚糖）在细胞壁分布的动态变化。在形成层及细胞伸展区域,细胞壁具有木葡聚糖的分布,而没有木聚糖和木质素沉积,随着次生壁S1层的形成,木质素出现在细胞角隅和胞间层,木聚糖开始出现在S1层中,此时木葡聚糖则分布在初生壁和胞间层;随着次生,壁S2层及S3层的形成和加厚,木质逐逐步由细胞角隅和胞间层扩展到S1、S2和S3层,其沉积呈现出不均匀的块状或片状沉积模式,在次生壁各层形成与其木质化的同时,木聚糖逐渐分布于整个次生壁中,而木糖聚糖仍局限分布于初生壁和胞间层。结果表明,随着细胞次生壁的形成与木质化,细胞壁结构发生较大变化。细胞壁的不同区域,如细胞角隅、胞间层、初生壁和次生壁各层,具有不同的半纤维素组成,其与木质等细胞壁组分结构构成不同的细胞壁分子结构。     

伯乐树茎次生木质部结构的研究

利用光镜和扫描电镜对伯乐树（Ｂｒｅｔｓｃｈｎｅｉｄｅｒａ ｓｉｎｅｎｓｉｓＨｅｍ ｓｌ．）茎次生木质部的结构进行了研究。其主要特征为：（１）散孔材,有较明显的生长轮;（２）导管分子多为单穿孔板,少数为梯形复穿孔板,具螺纹加厚;（３）管胞、纤维－管胞和韧型木纤维同时存在,后两者有的具分隔;（４）木薄壁组织以轮界分布为主;（５）木射线多为大型异形射线,属异形ＩＩＢ型;（６）缺乏侵填体、树脂道及分泌细胞。对伯乐树科（Ｂｒｅｔｓｃｈｎｅｉｄｅｒａｃｅａｅ）的系统位置作了探讨。     

不同纬度对鹅掌楸次生木质部结构的影响

对生长在北纬23至32度的鹅掌楸〖WTBX〗（Liriodendron chinense〖WTBZ〗 Sarg.）的次生木质部进行了比较解剖学研究。观察的特征有：导管分子的长度、直径、壁厚、梯状穿孔板横闩的数目;导管密度、纤维长度、直径、壁厚;射线的高度、宽度,单列射线的比例,射线的密度等。结果表明,随着纬度的增加,导管分子变短变细,壁变薄,梯状穿孔板的横闩数目减少;纤维也变短变细,壁变薄;木射线变矮变窄,单列射线的比例和射线密度增加。     

植物细胞程序死亡的机理及其与发育的关系

细胞程序死亡（PCD）是在植物体发育过程中普遍存在的,在发育的特定阶段发生的自然的细胞死亡过程,这一死亡过程是由某些特定基因编码的“死亡程序”控制的。PCD是细胞分化的最后阶段。细胞分化的临界期就处于死亡程序执行中的某个阶段。PCD包含启动期、效应期和清除期三个阶段,其间caspase家族起着重要作用。PCD在细胞和组织的平衡、特化,以及组织分化、器官建成和对病原体的反应等植物发育过程中起着重要作用。PCD中的形态学变化和生物化学变化都有着严格的时序性。植物的PCD和动物的PCD有许多共性,包括细胞形态和DNA降解等变化。也有一些不同,植物PCD的产物既可被其它细胞吸收利用;也可用于构建自身的次生细胞壁。     

Ultrastructural Changes in Cambial Cell Derivatives during Xylem Differentiation in Poplar

Abstract: The changes in cellular structures that occur in cambial cell derivatives during xylogenesis were examined in Populus trichocarpa Torr et Gray. During dormancy, the cells of the vascular cambium are characterised by dense cytoplasm, many small vacuoles and lipid bodies. During cambial activation, cambial cells are highly vacuolated, the cytoplasm is rich in organelles and the nucleus contains distinctly enlarged nucleoli. The plasma membrane forms vesicle-filled invaginations which mediate uptake of vesicular material into the vacuole. The mitotic patterns in dividing fusiform cells are fragmentary due to their strong vacuolisation. During cell enlargement, cambial cell derivatives remain strongly vacuolated and cytoplasmic structures are similar to active fusiform cells. From the beginning of secondary cell wall formation many changes in cytoplasmic structures occur in newly-formed fibres and vessels. In fibres, the cytoplasm is characterised by components of secondary cell wall synthesis, as indicated by increased amounts of endoplasmic reticulum, vesicle-producing dictyosomes and microtubules. In contrast, vessels show a more or less distinct occurrence of these components and remain more strongly vacuolated than fibres. Similar to cambial cells, a distinct flow of vesicular material into the vacuole through invaginations of the plasma membrane is apparent in fibres, as well as in vessels. After completion of the secondary cell walls, the loss of tonoplast integrity causes the collapse of the vacuole and initiates cell death in vessels and fibres. In vessels the tonoplast exhibits unusually strong staining prior to the collapse of the vacuole, indicating subsequent cell death. Overall, our results indicate an important role for the vacuole in the xylogen differentiation of cambial derivatives.     

Programmed Cell Death During the Vessel Element Differentiation of the Secondary Xylem in Eucommia ulmoides Shoots

The result from in situ end-labelling of fragmented DNA indicated that the vessel element differentiation of the secondary xylem in Eucommia ulmoides Oliv. was a typical programmed cell death (PCD) which involved a series of events, viz. synthesis of components essential for the secondary wall formation and a well organized succession of protoplast degeneration and autolysis in the tracheary cells. The nuclei gradually became irregular with highly condensed chromatin. In some nuclei, the cistema of the nuclear envelope became unevenly dilated within which some inner membrane protrusion enclosed with nuclear materials were present. The nuclear envelope underwent disruption and the nucleus eventually degenerated. However, as the nucleus was one of the most stable components in the cell, it was among the last organelles disappeared during the autolytic process. In the process, there were two forms of degeneration in the mitochondria (Mit). In one form the Mit shrank and became disorganized; in the other, part of the matrix in the Mit became electron-lucent with breakage of the membrane nearby. The cytoplasmic component residues were phagocitized and sequestered by the dilated rough endoplasmic reticulum (RER) cisternae. The RER and vacuoles did play a vital role in the further degeneration of other organelles just similar to the lysosomes acting in the animal cells. The autolyzed debri might be utilized in situ by taking part in the formation of secondary wall or be transported to the adjacent cells through the pits.     

天然型调节性T细胞发育分化的研究进展

CD4 CD25 调节性T细胞(Treg)对维持自身免疫耐受及调控免疫应答水平发挥非常重要的作用．Treg的缺失或紊乱导致如多发性硬化症、1型糖尿病等自身免疫性疾病的发生．研究发现,Treg在肿瘤免疫、感染免疫和移植免疫耐受中也发挥关键作用．根据来源不同,可将Treg分为胸腺来源的天然型Treg(natural Treg)和外周诱导型Treg(induced Treg)两群．天然型Treg(nTreg)是由胸腺发育分化成熟的,nTreg存在于胸腺CD4单阳性细胞中,表达CD4、CD25及叉头转录因子Foxp3,主要通过细胞与细胞之间直接接触发挥免疫抑制功能．研究表明,nTreg在胸腺中发育分化受到十分复杂的细胞、分子网络调控．胸腺微环境、T细胞受体、共刺激分子、IL-2等信号都可影响nTreg的发育分化．本文将主要对胸腺nTreg的发育分化过程及分子调控等方面进行综述．     

Mechanism of Xylem Differentiation in Pinus silvestrisL.

The duration and daily rate of radial growth and secondary-wallformation of all consecutive, radially-formed tracheids throughoutthe season were investigated in stems of adult trees of Pinussilvestris L. It was found that (1) the variation in radial diameter of tracheidswas probably dependent on seasonal changes in the rate of growthduring the phase of radial enlargement, (2) the daily rate ofcell-wall formation determined the final cell-wall thicknessof tracheids only at the beginning and the end of the season,(3) both rates were affected by temperature, (4) seasonal changesin cell-wall thickness were dependent mostly upon seasonal changesin the duration of the maturation period, (5) the changes inthe duration of the maturation period which brought about transitionfrom early to late wood were determined mostly by the delayin onset of autolysis of cytoplasm which terminates the phaseof tracheid maturation. This process, unlike the xylem productionfrom cambium and the termination of radial enlargemnt, was foundnot to be affected by the seasonal variation of temperature. An attempt to correlate these processes with the activity ofnatural auxin extracted from the cambial region gave negativeresults. On the basis of the results obtained, auxin and environmentalfactors such as precipitation and temperature seem not to bespecific for xylem differentiation. They may seriously affectwood differentiation if they become limiting or exceed the limitof tolerance, but probably they do not determine differentiationof the annual ring of conifers into early and late wood.     

Some Constituents of Xylem Sap and their Possible Relationship to Xylem Differentiation

Bleeding sap of Actinidia chinensis and Betula populifolia andguttation fluidof Avena sativa were analysed for sugars, amino-acids,auxin, and certain enzymes. A wide range of amino-acids wasfound in all three. Auxin was not detected in the bleeding sap,but was present in Avena guttation fluid (5.1 µg IAA equivalent/l).‘IAA oxidase’, acid phosphatase, ribonuclease, deoxyribonuclease,and protease were detected in the bleeding sap and guttationfluid. The possibility that some of the substances found insap and guttation fluid are products of autolysing, differentiatingxylem cells in the roots is discussed.