拟南芥种皮粘液质形成及调控机制研究进展

植物细胞壁是地球上储量最丰富的可再生资源,是人类生产和生活中能源、纤维、建筑材料和造纸等原料的主要来源。植物细胞壁的形成机制一直是近年来的研究热点,研究植物细胞壁的形成机制不仅有助于更高效地将细胞壁转化为生物乙醇等可再生能源,也将促进纤维生物质在食品、药品和纺织等领域的更高效利用,对于新能源开发和人类生产生活均具有十分重要的意义。一些十字花科(如拟南芥,Arabidopsis thaliana)和车前科植物的种皮外层细胞在发育过程中会合成和分泌大量的粘液质多糖,其在种子遇水后膨胀并释放,形成透明胶状物质包裹种子周围。拟南芥种皮粘液质的主要成分为果胶质(主要为鼠李半乳糖醛酸聚糖I),同时还含有少量的纤维素和半纤维素成分。种皮粘液质作为一种特化的细胞壁,具有表型容易观察、分离提取简便、组成相对单一、缺失不影响植株生长发育等优点,已成为研究植物细胞壁(果胶)多糖合成、调控及细胞壁组分间互作的理想模式体系,近年来取得了较大的研究进展,本文主要介绍拟南芥种皮粘液质的形成、组成及其调控机制方面的研究进展。     

多裂骆驼蓬种皮纹饰和粘液的解剖结构特征及其对种子扩散和萌发的影响

利用体视显微镜和扫描电子显微镜观察了荒漠植物多裂骆驼蓬粘液质种皮吸水前后的形态变化,通过种子吸水 脱水、粘土、漂浮和萌发实验研究了种皮纹饰和粘液特征在种子扩散和萌发中的适应意义,以期为研究该物种在荒漠环境中的适应策略提供理论依据。结果表明：（1）多裂骆驼蓬种皮纹饰由表皮细胞外切向壁的附属物向外突起形成,呈多面体网纹状;种皮纹饰上覆盖1层粘液薄膜,将多面体网眼围成封闭的腔,种子吸水后粘液薄膜变成凝胶状,腔内有气泡产生;粘液薄膜经过反复吸水 脱水后逐渐溶解消失。（2）种皮网眼状腔室结构和粘液薄膜可使种皮纹饰内储存空气,以增强种子的漂浮能力,有助于种子扩散;种皮纹饰和粘液还增强了种子的粘土能力,使种子锚定在土壤表面,避免种子裸露和活力丧失;此外,种皮纹饰及粘液的快速吸水和保水能力能防止种子失水,有效维持该物种在荒漠环境中的土壤种子库。（3）种皮纹饰和粘液虽可抑制种子萌发,但能促进幼根的伸长生长,对增强幼苗的建植能力有一定积极作用。     

新疆短命植物抱茎独行菜种子粘液质特性的研究

以新疆荒漠植物抱茎独行菜为材料,运用光镜与扫描电镜观察以及紫外吸收光谱法、化学反应及种子萌发实验等方法,对粘液质的形态和结构,物理化学特性,粘液质对种子萌发及萌发后的影响进行了研究.结果显示:(1)完整干种子表面覆盖着一层膜状物质(完全脱水的粘液质),并呈同一走向的山脊状突出的网状结构,遇水后粘液物质呈射线状向外发射出来,化学反应实验结果表明,粘液质的组成可能是某种多糖,如β-葡聚糖.(2)粘液质约占干种子重量的1/4,有很强的吸水能力,完全浸润10 min后,种子重量增加约30~40倍,种子长度、宽度、厚度的增加分别多于1倍、2倍、4倍;完全润湿的种子能够粘附相当于其干种子重量68倍的沙粒.(3)种皮粘液质对于不同土壤基质中的种子萌发有重要作用,但是对萌发后幼苗的生长没有作用.     

抱茎独行菜种皮粘液质相关基因TTG1的克隆、表达分析及功能鉴定

抱茎独行菜（Lepidium perfoliatum L.）为十字花科具典型粘液质繁殖体植物,而TTG1基因（Transpa-rent testa glabra 1）所编码的蛋白是调控种皮细胞分化并影响粘液质释放的转录因子。目前关于TTG1基因在粘液质繁殖体植物中的研究报道较少,为探究TTG1基因在抱茎独行菜粘液质发育中的作用,本研究利用同源克隆技术获得抱茎独行菜TTG1基因cDNA开放阅读框（ORF）序列,命名为LpTTG1。序列分析表明,该基因ORF全长为1032 bp,编码343个氨基酸,含有WD40基序;qRT-PCR分析结果显示,该基因在抱茎独行菜各组织中均有表达,反映了该基因功能的多样性;免疫组织化学定位结果表明,LpTTG1在种子发育过程中内珠被和外珠被的表达水平变化与外珠被粘液质的合成过程相一致,推测该基因可能参与调控抱茎独行菜种皮的发育及粘液质的形成。将LpTTG1基因转化拟南芥,该基因的过量表达显著促进了粘液质合成途径下游基因AtMUM4在角果中的表达,表明该基因有可能参与粘液质合成途径调控,并促进下游产物MUM4的产生。然而,对LpTTG1转基因拟南芥与野生型植株表型的比较发现,两者种子形态及粘液质分泌与释放方式均无显著差异,这可能是因为抱茎独行菜种皮发育和粘液质形成是一个多基因调控的复杂过程,某一基因的过量表达也许不会引起明显的表型变化。     

荒漠植物种子粘液的生态学意义

种子粘液是在种皮外层细胞的高尔基体内产生并分泌到胞腔内或细胞壁层的吸湿膨胀的一类果胶类多糖物质。具粘液种子的植物大多生长在荒漠地区,广泛存在于十字花科、菊科和车前科等类群中。粘液的存在对荒漠植物种子的扩散、萌发、防御以及幼苗的生长等都具有重要的生态学意义,是荒漠植物适应干旱少雨的生态环境的有效对策之一。对粘液种子的研究不仅可全面揭示荒漠植物的生态适应机制及其进化生态意义,还可为研究基因控制的糖类生物合成和分泌、细胞次生壁的生物合成及形态分化建立理想的模式体系。为此,在广泛查阅相关文献的基础上,该文综合分析了国内外种子粘液的研究进展,并重点探讨了以下几方面问题：（1）种子粘液的化学成分：（2）粘液及粘液种皮的形态特征：（3）粘液细胞分化与粘液生物合成的细胞学及基因调控机制以及粘液的释放方式：（4）种子粘液的生态学意义。在此基础上展望了今后的研究方向,以期为推动我国荒漠植物种子生态学的理论与应用研究及西部荒漠区的植物物种多样性保护和生态保育提供重要理论依据。     

荒漠植物种子粘液的生态学意义

种子粘液是在种皮外层细胞的高尔基体内产生并分泌到胞腔内或细胞壁层的吸湿膨胀的一类果胶类多糖物质。具粘液种子的植物大多生长在荒漠地区, 广泛存在于十字花科、菊科和车前科等类群中。粘液的存在对荒漠植物种子的扩散、萌发、防御以及幼苗的生长等都具有重要的生态学意义, 是荒漠植物适应干旱少雨的生态环境的有效对策之一。对粘液种子的研究不仅可全面揭示荒漠植物的生态适应机制及其进化生态意义, 还可为研究基因控制的糖类生物合成和分泌、细胞次生壁的生物合成及形态分化建立理想的模式体系。为此, 在广泛查阅相关文献的基础上, 该文综合分析了国内外种子粘液的研究进展, 并重点探讨了以下几方面问题: (1)种子粘液的化学成分; (2)粘液及粘液种皮的形态特征; (3)粘液细胞分化与粘液生物合成的细胞学及基因调控机制以及粘液的释放方式; (4)种子粘液的生态学意义。在此基础上展望了今后的研究方向, 以期为推动我国荒漠植物种子生态学的理论与应用研究及西部荒漠区的植物物种多样性保护和生态保育提供重要理论依据。     

桃儿七种子休眠解除过程中细胞壁代谢及种皮超微结构的变化北大核心CSCD

为探究低温层积过程中桃儿七种子细胞壁代谢及种皮超微结构与休眠解除的内在联系,该研究通过低温层积解除桃儿七种子休眠,分析休眠解除过程中种子不同部位细胞壁组分及相关代谢酶的变化,同时利用扫描电镜对种皮的超微结构进行观察。结果表明,(1)桃儿七种皮主要由角质层、栅状石细胞层及海绵组织层3层构成,在层积过程中,种皮内部的海绵组织逐步疏松膨胀,种皮表面破损加剧;(2)种子不同部位的细胞壁组分具有明显差异,整个层积过程中,种胚、种皮和胚乳中的纤维素含量均在层积中期(45 d和60 d)降至最低,3个部位的纤维素酶活性在层积中期对应升高;种胚和种皮内的半纤维素含量均在层积中期显著下降,种皮中甘露聚糖酶活性和木糖苷酶活性在层积中期时相应达到最大;3个部位的果胶含量均在层积后期(75 d和90 d)时显著下降,而种皮和胚乳中多聚半乳糖醛缩酶活性也在层积后期相应升高;(3)种胚和胚乳内过氧化物酶活性在层积75 d和90 d时明显下降,而SOD活性在此时显著上升。(4)种子不同部位3种木质素单体的组成比例具有明显区别,同时3种木质素单体含量均随层积时间的延长而显著降低,且胚乳和种皮中的S-木质素含量对种子萌发存在显著的负向影响关系。研究认为,在低温层积过程中,桃儿七种子内细胞壁组分纤维素、半纤维素及木质素的逐步酶解,活性氧作用下的细胞壁松弛以及海绵组织层的疏松膨胀和种皮的破裂,破坏了细胞壁的刚性结构,促使种子机械束缚力降低,吸水性能提高、胚根生长能力增强,最终导致其休眠解除。     

朱唇种子吸水特性及其在干旱胁迫下萌发特性

种子粘液质是植物在长期适应环境过程中形成的,该物质对于种子的扩散、定居、生存力的改善、萌发、幼苗生存乃至抵御有毒化学物质毒害等都具有重要的生态学意义。朱唇为唇形科鼠尾草属多年生草本植物,原产美洲热带地区,现已广泛栽植于世界各地。为了理解朱唇种子表面的粘液物质吸水特性和种子在干旱胁迫下的萌发特性,该研究以朱唇种子为材料,运用光学显微镜和扫描电子显微镜观察以及种子萌发试验的方法,对种子和粘液层的形态结构、粘液质对种子萌发的影响进行了研究。结果表明:朱唇种子为卵形,表面为负网状结构,千粒重为(1.611±0.0084)g,无粘液种子吸水倍数为3,粘液种子吸水倍数为25,粘液层吸水倍数为122。粘液和无粘液种子及粘液层的重量都随吸水时间的延长而增长,但脱水过程要远长于吸水过程。朱唇种子吸水2 h达到饱和,经过36 h可干燥失水恢复原重。不同浓度PEG对朱唇种子的萌发均有影响,发芽势随PEG浓度升高而显著降低。朱唇种子在5%PEG胁迫下发芽率最高达(90.00±8.66)%,20%PEG胁迫下发芽率最低为(76.67±10.41)%,低浓度PEG对朱唇种子萌发有一定促进作用。这说明朱唇种子为速萌型种子,其粘液质在种子吸水过程中起到举足轻重的作用,能保证短时间内有充足的水分供其萌发。     

兰花蕉种子的解剖学和组织学研究

兰花蕉种子球形成或近球形,具表皮毛,种脊不明显,种子包括假种皮,种皮,外胚乳,内胚乳和胚五部分,假种皮具３～４条粗毛状裂片,包围种子或不定向伸展,裂片最外方为１层表皮细胞和１～３层厚壁细胞,内方为薄壁细胞;表皮细胞和厚壁细胞的壁增厚并木质化,成熟时裂片下部１／２段中空,种皮由外珠被发育而来,但内珠被在种子发育后期才萎缩,种皮分化为外种皮,中种皮与内种皮;外种皮由１层有皮细胞构成,其细胞壁增厚并木质     

苏铁种子的种皮结构特征研究

对苏铁(Cycas revoluta Thunb.)种子的种皮进行了解剖研究,结果表明:苏铁种子的种皮分为外种皮、中种皮和内种皮3层结构.外种皮含有角质化的表皮细胞、薄壁细胞以及少量的厚壁细胞和异细胞,布有树脂道、气室和4束大维管束;中种皮主要由厚壁细胞群和木质化纤维组成,种孔端有一条缝合线,种脐端有3个孔;内种皮由多层干瘪的薄壁细胞和脉络状维管束组成,种孔端有一层椭圆状保护膜.对外种皮和内种皮维管束进行观察研究发现:外种皮和内种皮的维管束分布方式及其结构存在明显差异,外种皮的维管束由种脐端顺着种子弧形走向种孔端,内种皮的维管束呈脉络状,形成维管网贯穿其中;内、外种皮维管束中均存在多种不同样式的导管.     

A subtilisin-like serine protease essential for mucilage release from Arabidopsis seed coats

During Arabidopsis seed development large quantities of mucilage, composed of pectins, are deposited into the apoplast underneath the outer wall of the seed coat. Upon imbibition of mature seeds, the stored mucilage expands through hydration and breaks the outer cell wall that encapsulates the whole seed. Mutant seeds carrying loss-of-function alleles of AtSBT1.7 that encodes one of 56 Arabidopsis thaliana subtilisin-like serine proteases (subtilases) do not release mucilage upon hydration. Microscopic analysis of the mutant seed coat revealed no visible structural differences compared with wild-type seeds. Weakening of the outer primary wall using cation chelators triggered mucilage release from the seed coats of mutants. However, in contrast to mature wild-type seeds, the mutant's outer cell walls did not rupture at the radial walls of the seed coat epidermal cells, but instead opened at the chalazal end of the seed, and were released in one piece. In atsbt1.7, the total rhamnose and galacturonic acid contents, representing the backbone of mucilage, remained unchanged compared with wild-type seeds. Thus, extrusion and solubility, but not the initial deposition of mucilage, are affected in atsbt1.7 mutants. AtSBT1.7 is localized in the developing seed coat, indicating a role in testa development or maturation. The altered mode of rupture of the outer seed coat wall and mucilage release indicate that AtSBT1.7 triggers the accumulation, and/or activation, of cell wall modifying enzymes necessary either for the loosening of the outer primary cell wall, or to facilitate swelling of the mucilage, as indicated by elevated pectin methylesterase activity in developing atsbt1.7 mutant seeds.     

Cellulose synthesis via the FEI2 RLK/SOS5 pathway and cellulose synthase 5 is required for the structure of seed coat mucilage in Arabidopsis

The seeds of Arabidopsis thaliana and many other plants are surrounded by a pectinaceous mucilage that aids in seed hydration and germination. Mucilage is synthesized during seed development within maternally derived seed coat mucilage secretory cells (MSCs), and is released to surround the seed upon imbibition. The FEI1/FEI2 receptor-like kinases and the SOS5 extracellular GPI-anchored protein were shown previously to act on a pathway that regulates the synthesis of cellulose in Arabidopsis roots. Here, we demonstrate that both FEI2 and SOS5 also play a role in the synthesis of seed mucilage. Disruption of FEI2 or SOS5 leads to a reduction in the rays of cellulose observed across the seed mucilage inner layer, which alters the structure of the mucilage in response to hydration. Mutations in CESA5, which disrupts an isoform of cellulose synthase involved in primary cell wall synthesis, result in a similar seed mucilage phenotype. The data indicate that CESA5-derived cellulose plays an important role in the synthesis and structure of seed coat mucilage and that the FEI2/SOS5 pathway plays a role in the regulation of cellulose synthesis in MSCs. Moreover, these results establish a novel structural role for cellulose in anchoring the pectic component of seed coat mucilage to the seed surface.     

The role of COBRA‐LIKE 2 function,as part of the complex network of interacting pathways regulating Arabidopsis seed mucilage polysaccharide matrix organization

The production of hydrophilic mucilage along the course of seed coat epidermal cell differentiation is a common adaptation in angiosperms. Previous studies have identified COBRA‐LIKE 2 (COBL2), a member of the COBRA‐LIKE gene family, as a novel component required for crystalline cellulose deposition in seed coat epidermal cells. In recent years, Arabidopsis seed coat epidermal cells (SCEs), also called mucilage secretory cells, have emerged as a powerful model system for the study of plant cell wall components biosynthesis, secretion, assembly and de muro modification. Despite accumulating data, the molecular mechanism of COBL function remains largely unknown. In the current research, we utilized genetic interactions to study the role of COBL2 as part of the protein network required for seed mucilage production. Using correlative phenotyping of structural and biochemical characteristics, unique features of the cobl2 extruded mucilage are revealed, including: ‘unraveled’ ray morphology, loss of primary cell wall ‘pyramidal’ organization, reduced Ruthenium red staining intensity of the adherent mucilage layer, and increased levels of the monosaccharides arabinose and galactose. Examination of the cobl2cesa5 double mutant provides insight into the interface between COBL function and cellulose deposition. Additionally, genetic interactions between cobl2 and fei1fei2 as well as between each of these mutants to mucilage‐modified 2 (mum2) suggest that COBL2 functions independently of the FEI‐SOS pathway. Altogether, the presented data place COBL2 within the complex protein network required for cell wall deposition in the context of seed mucilage and introduce new methodology expending the seed mucilage phenotyping toolbox.     

Understanding polysaccharide production and properties using seed coat mutants: future perspectives for the exploitation of natural variants

Background

The epidermal cells of the seed coat of certain species accumulate polysaccharides during seed development for cell wall reinforcement or release on imbibition to form mucilage. Seed-coat epidermal cells show natural variation in their structure and mucilage production, which could explain the diverse ecophysiological roles proposed for the latter. Arabidopsis mucilage mutants have proved to be an important tool for the identification of genes involved in the production of seed-coat polysaccharides.

Scope

This review documents genes that have been characterized as playing a role in the differentiation of the epidermal cells of the arabidopsis seed coat, the natural variability in polysaccharide features of these cells and the physiological roles attributed to seed mucilage.

Conclusions

Seed-coat epidermal cells are an excellent model for the study of polysaccharide metabolism and properties. Intra- and interspecies natural variation in the differentiation of these epidermal cells is an under-exploited resource for such studies and promises to play an important part in improving our knowledge of polysaccharide production and ecophysiological function.     

Differentiation of mucilage secretory cells of the Arabidopsis seed coat

In some plant species, including Arabidopsis, fertilization induces the epidermal cells of the outer ovule integument to differentiate into a specialized seed coat cell type with a unique morphology and containing large quantities of polysaccharide mucilage (pectin). Such seed coat mucilage cells are necessary for neither viability nor germination under normal laboratory conditions. Thus, the Arabidopsis seed coat offers a unique system with which to use genetics to identify genes controlling cell morphogenesis and complex polysaccharide biosynthesis and secretion. As a first step in the application of this system, we have used microscopy to investigate the structure and differentiation of Arabidopsis seed coat mucilage cells, including cell morphogenesis and the synthesis, secretion, and extrusion of mucilage. During seed coat development in Arabidopsis, the epidermal cells of the outer ovule integument grow and differentiate into cells that produce large quantities of mucilage between the primary cell wall and plasma membrane. Concurrent with mucilage production, the cytoplasm is shaped into a column in the center of the cell. Following mucilage secretion the cytoplasmic column is surrounded by a secondary cell wall to form a structure known as the columella. Thus, differentiation of the seed coat mucilage cells involves a highly regulated series of events including growth, morphogenesis, mucilage biosynthesis and secretion, and secondary cell wall synthesis.     

The FEI2-SOS5 pathway and CELLULOSE SYNTHASE 5 are required for cellulose biosynthesis in the Arabidopsis seed coat and affect pectin mucilage structure

A common adaptation in angiosperms is the deposition of hydrophilic mucilage into the apoplast of seed coat epidermal cells during the course of their differentiation. Upon imbibition, seed mucilage, composed mainly of carbohydrates (i.e. pectins, hemicelluloses and glycans) expands rapidly, encapsulating the seed and aiding in seed dispersal and germination. The FEI1/FEI2 receptor-like kinases and the SOS5 extracellular GPI-anchored protein were previously shown to act on a pathway regulating cellulose biosynthesis during Arabidopsis root elongation. In the highlighted study, we demonstrated that FEI2 and SOS5 regulate the production of the cellulosic rays deposited across the inner adherent-layer of seed mucilage. Mutations in either fei2 or sos5 disrupted the formation of rays, which was associated with an increase in the soluble, outer layer of pectin mucilage and accompanied by a reduction in the inner adherent-layer. Mutations in CELLULOSE SYNTHASE 5 also led to reduced rays and mal-partitioning of the pectic component of seed mucilage, further establishing a structural role for cellulose in seed mucilage. Here, we show that FEI2 expressed from a CaMV 35S promoter complemented both root and seed mucilage defects of the fei1 fei2 double mutant. In contrast, expression of FEI1 from a 35S promoter complemented the root, but not the seed phenotype of the fei1 fei2 double mutant, suggesting that unlike in the root, FEI2 plays a unique and non-redundant role in the regulation of cellulose synthesis in seed mucilage. Altogether, these data suggest a novel role for cellulose in anchoring the pectic component of seed mucilage to the seed surface and indicate that the FEI2 protein has a function distinct from that of FEI1, despite the high sequence similarity of these RLKs.     

Ontogeny of solasodine-containing mucilage layer inSolanum viarum Dunal, ploidy types

Berries of steroid-bearingSolanum viarum Dunal are exploited commercially in India as raw material by steroid industries for solasodine, a glycoalkaloid, present in the mucilaginous exotesta of the seed. Comparative ontogeny of exotesta studied through histochemical studies in diploid, autotetraploid and trisomic plants indicated similarity in the histochemical changes occurring during ontogeny of the outermost seed coat layer which culminated in the transformation of this layer into the mucilage layer. The increased cell size in this layer in the autotetraploid plants probably accounts for the higher steroid content reported. Corroborative evidences for histochemical changes observed in the mucilage layer were obtained from studies of ultrastructure using transmission electron microscopy.