河西走廊芦苇的光合碳同化途径对生境条件的适应

本文以甘肃省河西走廊生长的四个不同生境芦苇为对象,比较研究了它们的叶解剖结构,光合关键酶活力,乙醇酸氧化酶活力和稳定碳同位素组成（δ13C）,结果发现,沼泽芦苇叶中虽具有不典型的花环结构,但维管束鞘细胞中不含叶绿体,RUBPcase活力／PEPcase活力比值为24．4,乙醇酸氧化活力为1218Unit mgpro^-1.min^-1δ13C值为－34‰,这些测值位于C3植物（小麦）的范围内,生长于沙兵上的芦苇叶片具有明显的花环结构,维管束鞘细胞内含异型叶绿体,RUBPcase活力／PEPcase活力比值为0．985,乙醇酸氧化酶活力为504Unit mg pro^-1.min^-1,δ13C值为－20．9％,这些测值与典型C4植物玉米十分相似。盐化草甸芦苇和盐化草－－沙丘过渡地带芦苇叶中均具有明显的花环结构,维管束鞘细胞中含大型叶绿体,RUBPcase活力／PEPcase 活力比值分别为2．45和1．53,但乙醇酸氧化酶活力分别为1470和2058Unmitmgpro^-1,min^-1,δC值分别为35．6和30．6‰综合盐化草甸芦苇和过渡地带芦苇的上述指标,似介于沼泽芦苇和沙丘芦苇之间,由这些结果可以认为,分布于甘肃省河西走廊的芦苇,在种内发生有由环境因子引起的光合碳代谢途径的适应性改变。     

四种生态型芦苇叶片结构的研究

采用光学显微镜和电子显微镜对４种生态型芦苇（ＰｈｒａｇｍｉｔｅｓｃｏｍｍｕｎｉｓＴｒｉｎ．）叶解剖结构及叶脉维管束鞘细胞超微结构进行了观察研究。结果表明,沼泽芦苇维管束鞘细胞内含物较少,维管束鞘细胞叶绿体呈近圆形,超微结构与典型Ｃ３植物小麦鞘细胞的相似,基粒发育退化,不含淀粉粒;沙丘芦苇鞘细胞内含物较多,鞘细胞叶绿体呈近椭圆形,超微结构与典型Ｃ４植物玉米的相似,具有不发达的基粒;重度盐化草甸芦苇鞘细胞内含物则较沼泽芦苇多,而较沙丘芦苇少,鞘细胞叶绿体形态、超微结构与沙丘芦苇相似;轻度盐化草甸芦苇鞘细胞内含物与沙丘芦苇相似,但是,其鞘细胞含有与沼泽芦苇鞘细胞内叶绿体形态相似的近圆形以及与沙丘芦苇和重度盐化草甸芦苇鞘细胞内形态相似的近椭圆形两种形态的叶绿体。表明生长在同一地区不同环境中的４种生态型芦苇的光合结构发生了深刻的变异,表现出从Ｃ３向Ｃ４进化的明显趋势。     

芦苇耐脱水能力的生理生态学分析

甘肃省河西走廊分布有不同生态环境的芦苇,包括沼泽芦苇、盐化草甸芦苇和沙丘芦苇。盐化草甸芦苇和沙丘芦苇表观出很强的耐脱水能力。 测定与植物耐脱水能力相关的SOD、CAT和POD活力,表现为SOD活力以沙丘芦苇最高,盐化草甸芦苇次之,沼泽芦苇最低,但CAT和POD则以沼泽芦苇高于其它生境芦苇。用聚丙烯酰胺垂直板电泳方法分析这3种酶同工酶谱,结果表明,3种不同生境芦苇的SOD同工酶谱基本相同,均有9条酶带,但沙丘芦苇和盐化草甸芦苇的CAT和POD同工酶带数比沼泽芦苇分别增多3条和2条。这些结果证明,芦苇在自然选择压力下形成的耐脱水能力与基因调控的酶多型性相关。     

四种生态型芦苇叶中离子分布对生境的生理适应

采用X射线微区分析技术,测定了4种生态型芦苇(Phragmites australis (CaV.) Trin. exSteud.)叶的表皮泡状细胞、叶肉细胞和叶脉维管束鞘细胞离子的含量.结果表明:沼泽芦苇的鞘细胞内,K+、Na+、Ca2+、Mg2+和Cl-分布均较叶肉细胞和泡状细胞高.沙丘芦苇的泡状细胞中Ca2+分布较叶肉细胞和鞘细胞高,而Mg2+在其叶肉细胞,以及K+、Na+和Cl-在其鞘细胞内分布均较高.在轻度盐化草甸芦苇的叶肉细胞内分布较多的Na+和Mg2+,而在鞘细胞内K+、Ca2+ 和Cl-的分布均较叶肉细胞和泡状细胞为高.重度盐化草甸芦苇的泡状细胞内Na+和Mg2+的分布较多;同样,在叶肉细胞中K+、Ca2+和Cl-的分布也较多.最后,讨论了上述各种离子在不同生态型芦苇叶内分布的状况, 以及与其环境适应的生理意义.     

四种生态型芦苇叶中离子分布对生境的生理适应

采用X射线微区分析技术 ,测定了 4种生态型芦苇 (Phragmitesaustralis (CaV .)Trin .exSteud .)叶的表皮泡状细胞、叶肉细胞和叶脉维管束鞘细胞离子的含量。结果表明 :沼泽芦苇的鞘细胞内 ,K 、Na 、Ca2 、Mg2 和Cl-分布均较叶肉细胞和泡状细胞高。沙丘芦苇的泡状细胞中Ca2 分布较叶肉细胞和鞘细胞高 ,而Mg2 在其叶肉细胞 ,以及K 、Na 和Cl- 在其鞘细胞内分布均较高。在轻度盐化草甸芦苇的叶肉细胞内分布较多的Na 和Mg2 ,而在鞘细胞内K 、Ca2 和Cl- 的分布均较叶肉细胞和泡状细胞为高。重度盐化草甸芦苇的泡状细胞内Na 和Mg2 的分布较多 ;同样 ,在叶肉细胞中K 、Ca2 和Cl- 的分布也较多。最后 ,讨论了上述各种离子在不同生态型芦苇叶内分布的状况 ,以及与其环境适应的生理意义。     

两种生物型芦苇胚性悬浮培养物对渗透胁迫的生理响应 Ⅱ．抗氧化酶类活性的变化

渗透胁迫处理下,沙丘芦苇和沼泽芦苇培养细胞的H2O2含量变化不大,MDA含量随时间延长而降低。抗氧化酶中,沙丘芦苇的SOD、CAT、POD和APx的活力均显著高于沼津芦苇,显示出比沼泽芦苇更强的ROS清除能力。胁迫初期（0．5h-1d)沙丘芦苇细胞中主要表现出SOD、CAT活力的提高,胁迫2d后4种酶活力均明显被诱导。两种生态型芦苇的PEG适应性培养物APx及SOD酶活力显著提高,POD活性仅沼泽芦苇中升高。提示SOD、CAT主要参与沙丘芦苇在渗透胁迫下的短期响应,而APx与长期响应有关。     

河西走廊不同生态型芦苇核酸代谢季节动态研究

分布在甘肃河西走廊的４ 种生态型芦苇（Ｐｈｒａｇｍ ｉｔｅｓｃｏｍ ｍ ｕｎｉｓＴｒｉｎ．）的核酸代谢季节变化有差异。盐化草甸芦苇ＲＮＡ 含量持续增加,ＤＮＡ 含量相对稳定,其它３ 种生态型芦苇的ＲＮＡ 和ＤＮＡ 含量以５月份为最高。过渡带芦苇的ＲＮＡ 含量、沼泽芦苇及沙丘芦苇的ＤＮＡ含量９ 月份略有增高。盐化草甸芦苇与过渡带芦苇的ＤＮａｓｅ和ＲＮａｓｅ的活性７ 月份最高,沼泽芦苇与沙丘芦苇的ＤＮａｓｅ和ＲＮａｓｅ活性５—９ 月份呈增高趋势。盐化草甸芦苇的ＤＮＡ 和ＲＮＡ 合成活性不断升高,过渡带芦苇和沼泽芦苇的ＤＮＡ 和ＲＮＡ 合成活性及沙丘芦苇的ＲＮＡ 合成活性５—９月份均降低,仅沙丘芦苇的ＤＮＡ合成活性增强。ＲＮＡ 聚丙烯酰胺梯度凝胶电泳分析结果表明,４ 种生态型芦苇均含有２５Ｓ、２３Ｓ、１８Ｓ、１６Ｓ大分子量ｒＲＮＡ 和小分子量５．８Ｓ、５Ｓ、４．５ＳｒＲＮＡ 及４ＳｔＲＮＡ。大分子量ＲＮＡ 的含量高于小分子量ＲＮＡ 含量。不同生态型及同一生态型芦苇的不同发育时期,相同的ＲＮＡ 组分含量各不相同。且发育过程中２３Ｓ、１８Ｓ、１６ＳｒＲＮＡ 在不同月份发生不同程度的降解。由此,我们认为,核酸代谢的差异性是４ 种生态型由生长转入衰     

芦苇生态型划分指标的主分量及模糊聚类分析

运用主分量分析及模糊聚类的方法首次对包括群落、形态,解剖,生理生化及遗传代谢等４５个指标的不同生态芦苇进行了分析,结果表明,生物量、单株干重、株高,可溶性蛋白质含量、ＲＮＡ含量、ＤＮａａｓｅ和ＲＮａｓｅ活性、Ｋｒａｎｚ结构的有无及土壤ｐＨ值等差异最为显著。４种生态型芦苇中,盐化草甸芦苇与盐化草甸－沙丘过渡带芦苇最相近,其次为沙丘芦苇,沼泽芦苇与其它３种生态相似程度最低。     

若尔盖草甸退化对土壤碳、氮和碳稳定同位素的影响

为了解不同退化阶段高寒草甸土壤碳、氮和碳稳定同位素的差异,对若尔盖湿地内沼泽草甸、草原化草甸、退化草甸3个阶段土壤的碳、氮和碳稳定同位素进行了分析.结果表明:若尔盖湿地草甸土壤δ¹³C 值介于-26.21‰～-24.72‰之间,土壤δ¹³C 值随土层加深而增大.土壤δ¹³C 值与有机碳含量对数值呈线性负相关.表层土壤(0～10 cm)δ¹³C值大小顺序为草原化草甸>退化草甸>沼泽草甸,β值大小顺序为草原化草甸>沼泽草甸>退化草甸.沼泽草甸、草原化草甸、退化草甸0～30 cm 土壤碳含量分别为105.32、42.11和31.12 g·kg^-1,氮含量分别为8.74、3.41和2.81 g·kg^-1,C/N分别为11.26、11.23和10.89.随着草甸的退化,土壤碳、氮呈降低趋势,退化草甸C/N值低于沼泽草甸和草原化草甸.随着土层深度加深,碳、氮含量呈现降低趋势.草甸退化导致的土壤δ¹³C 值差异主要发生在表层0～10 cm.3个退化阶段中,退化草甸土壤的β值和C/N最低,表明退化草甸土壤矿化作用较强.     

两种生态型芦苇胚性悬浮培养物对渗透胁迫的生理响应Ⅱ.抗氧化酶类活性的变化

渗透胁迫处理下,沙丘芦苇和沼泽芦苇培养细胞的H2O2含量变化不大,MDA含量随时间延长而降低.抗氧化酶中,沙丘芦苇的SOD、CAT、POD和APx的活力均显著高于沼泽芦苇,显示出比沼泽芦苇更强的ROS清除能力.胁迫初期(O.5 h～1 d)沙丘芦苇细胞中主要表现出SOD、CAT活力的提高,胁迫2 d后4种酶活力均明显被诱导.两种生态型芦苇的PEG适应性培养物APx及SOD酶活力显著提高,POD活性仅沼泽芦苇中升高.提示SOD、CAT主要参与沙丘芦苇在渗透胁迫下的短期响应,而APx与长期响应有关.     

四种生态型芦苇叶片结构的研究

The leaf anatomy and uhrastructure of the vascular bundle sheath cells of four ecotypes of reed (Phragmites communis Trin. ) were observed with light and electron microscope. The four ecotypes were the swamp reed, dune reed, heavy salt meadow reed and the light salt meadow reed. It was shown that the intracellular content of the vascular bundle sheath cells of the swamp reed was lower than that of the dune reed; that of the heavy salt meadow reed was between that of the former two reeds and that of the light salt meadow reed and that of the light salt meadow reed was similar to the dune reed. The chloroplasts in the swamp reed were round-shaped with poorly developed grana. Starch grains were not visualized. The uhrastructure of their chloroplasts were similar to those in the typical C3 plants as wheat. By contrast, the chloroplasts in the vascular bundle cells of the dune reed were elliptical with their structure similar to that of the typical C4 plants as maize. These chloroplasts contained some poorly developed grana thykloid. The chloroplasts of the vascular bundle sheath cells in the heavy salt meadow reed were similar to those seen in the dune reed, however, those in the light salt meadow reed appeared to be beth round .and elliptical in shape. The above results suggested a great habitat variation did occur among the four ecotypes of reed in which these exists an obvious evolutional trend from C3 to C4 plants.     

The Identification of Photosynthetic Subpathways of Some C4 and CAM Plants

The photosynthetic subpathways of five C4 plants and one CAM plant were distinguished according to their chemical, physiological and cytological characteristics. Based on C4 acid decarboxylation enzymes, four C4 plants of Setaria glauca, Sporobolus indicus, Zoysia tenuifolia and Leptochloa chinensis all exhibited the functional high activities of PEP carboxykinase and aspartate aminotransferase as seen in the known PEP-CK subtype. The δ13C value of –12.43% in leaves of L. chinensis was also consistent with that range among PEP-CK subtype. So, these species were classified into PEP-CK subtype. However, their chloroplasts in bundle sheath cells were evenly distributed, not as that displayed centrifugally or centripetally in three typical subtypes. The even arrangement of chloroplasts in bundle sheath cells was likely to be an evolutional intermediate from centripetal (NAD ME type) to centrifugal types (NADP-ME and most PEP-CK types). The high activities of NAD-malic enzyme and aspartate aminotransferase, accompanied with the centripetally located chloroplasts, 0.057 of quantum yield and tile δ13C value of –15.3% in leaves of C4 dicot Euphobia hirta indicated characteristics of NAD-ME subtype. Moreover, CAM plant Aloe vera clearly fell into PEP-CK sybtype because of its high activity of PEP-CK both in whole leaf and green tissue.     

A survey of photosynthetic carbon metabolism in 4 ecotypes of Phragmites australis in northwest China: Leaf anatomy, ultrastructure, and activities of ribulose 1,5-bisphosphate carboxylase, phosphoenolpyruvate carboxylase and glycollate oxidase

Four ecotypes of Phragmites australis from different habitats in northwest China were examined to compare their photosynthetic characteristics. In a swamp ecotype, the Δ ¹³C value of leaf materials was −34.0‰, and bundle sheath cells contained a small amount of organelles and round-shaped chloroplasts, as being similar to typical C₃ plants. In a dune ecotype, the Δ ¹³C value was −20.9‰ and bundle sheath cells contained oval-shaped chloroplasts with poorly-developed grana. In light and heavy salt meadow ecotypes, Δ ¹³C values were −30.6‰ and −35.6‰, respectively. The shape of bundle sheath chloroplasts in the light salt meadow ecotype was intermediate between those of the swamp and dune ecotypes. Abundance of bundle sheath organelles in the heavy salt meadow ecotype was intermediate. The swamp ecotype had photosynthetic enzyme activities typical of C₃ type plants, whereas the dune ecotype had an increased activity of phosphoenolpyruvate carboxylase (PEPC), a key C₄ enzyme, and a decreased ribulose 1,5-bisphosphate carboxylase (Rubisco) activity. The light salt meadow and heavy salt meadow ecotypes had substantial activities of PEPC, which indicates potential for C₄ photosynthesis. These data suggest that this species evolved the C₃-like ecotype in swamp environments and the C₄-like C₃-C₄ intermediate in dune desert environments, and C₃-like C₃-C₄ intermediates in salt environments.     

Leaf anatomy and C4 photosynthetic enzymes in three reed ecotypes

Differences in leaf interveinal distances, chloroplasts distribution in bundle sheath cells (BSC) and activities of C₄ photosynthetic enzymes in the leaves of three ecotypes of Phragmites communis Trinius, namely swamp reed (SR), heavy salt meadow reed (HSMR) and dune reed (DR), occurring in the desert region of northwest China were investigated. The two terrestrial ecotypes, DR and HSMR, had denser vascular system, more and longer BSC chloroplasts and higher capacity of CO₂ concentrating mechanism of NAD-ME subtype as compared with the SR ecotype. The enhanced NADP-ME pathway in the HSMR might contribute to its adaptation to the salinity habitat.     

Anatomy, chloroplast structure and compartmentation of enzymes relative to photosynthetic mechanisms in leaves and cotyledons of species in the tribe Salsoleae (Chenopodiaceae)

Certain members of the family Chenopodiaceae are the dominant species of the deserts of Central Asia; many of them are succulent halophytes which exhibit C4-type CO2 fixation of the NAD- or NADP-ME (malic enzyme) subgroup. In four C4 species of the tribe Salsoleae, the Salsoloid-type Kranz anatomy in leaves or stems was studied in relation to the diversity in anatomy which was found in cotyledons. Halocharis gossypina, has C4 NAD-ME Salsoloid-type photosynthesis in leaves and C3 photosynthesis in dorsoventral non-Kranz cotyledons; Salsola laricina has C4 NAD-ME Salsoloid-type leaves and C4 NAD-ME Atriplicoid-type cotyledons; Haloxylon persicum, has C4 NADP-ME Salsoloid-type green stems and C3 isopalisade non-Kranz cotyledons; and S. richteri has C4 NADP-ME Salsoloid-type leaves and cotyledons. Immunolocalization studies on Rubisco showed strong labelling in bundle sheath cells of leaves and cotyledons of organs having Kranz anatomy. The C4 pathway enzyme phosphoenolpyruvate carboxylase was localized in mesophyll cells, while the malic enzymes were localized in bundle sheath cells of Kranz-type tissue. Immunolocalization by electron microscopy showed NAD-ME is in mitochondria while NADP-ME is in chloroplasts of bundle sheath cells in the respective C4 types. In some C4 organs, it was apparent that subepidermal cells and water storage cells also contain some chloroplasts which have Rubisco, store starch, and thus perform C3 photosynthesis. In non-Kranz cotyledons of Halocharis gossypina and Haloxylon persicum, Rubisco was found in chloroplasts of both palisade and spongy mesophyll cells with the heaviest labelling in the layers of palisade cells, whereas C4 pathway proteins were low or undetectable. The pattern of starch accumulation correlated with the localization of Rubisco, being highest in the bundle sheath cells and lowest in the mesophyll cells of organs having Kranz anatomy. In NAD-ME-type Kranz organs (leaves and cotyledons of S. laricina and leaves of H. gossypina the granal index (length of appressed membranes as a percentage of total length of all membranes) of bundle sheath chloroplasts is 1.5 to 2.5 times higher than that of mesophyll chloroplasts. In contrast, in the NADP-ME-type Kranz organs (S. richteri leaves and cotyledons and H. persicum stems) the granal index of mesophyll chloroplasts is 1.5 to 2.2 times that of the bundle sheath chloroplasts. The mechanism of photosynthesis in these species is discussed in relation to structural differences.     

Ultrastructure of leaves in C4 Cyperus iria and C3 Carex siderosticta

The ultrastructural aspects ofCyperus iria leaves showing the C₄ syndrome and the typical C₃ species,Carex siderosticta, in the Cyperaceae family were examined.C. iria exhibited the chlorocyperoid type, showing an unusual Kranz structure with vascular bundles completely surrounded by two bundle sheaths. The cellular components of the inner Kranz bundle sheath cells were similar to those found in the NADP-ME C₄ subtype, having centrifugally arranged chloroplasts with greatly reduced grana and numerous starch grains. Their chloroplasts contained convoluted thyla-koids and a weakly-developed peripheral reticulum, although it was extensive mostly in mesophyll cell chloroplasts. The outer mestome bundle sheath layer was sclerenchymatous and generally devoid of organelles, but had unevenly thickened walls. Suberized lamellae were present on its cell walls, and they became polylamellate when traversed by plasmodesmata. Mesophyll cell chloroplasts showed well-stacked grana with small starch grains. InC. siderosticta, vascular bundles were surrounded by the inner mestome sheath and the outer parenchymatous bundle sheath with intercellular spaces. The mestome sheath cells degraded in their early development and remained in a collapsed state, although the suberized lamellae retained polylamellate features. Plastids with a crystalline structure, sometimes membrane-bounded, were found in the epidermal cells. The close interveinal distance was 35–50 μm inC. iria, whereas it was 157–218 μm inC. siderosticta. These ultrastructural characteristics were discussed in relation to their photosynthetic functions.     

The occurrence of C4 species in the genusRhynchospora and its significance in kranz anatomy of the cyperaceae

Rhynchospora rubra was found to have a low CO₂ compensation point, high δ¹³C value, Kranz leaf anatomy, starch present in the bundle sheath cells and narrow interveinal distance. These observations suggest thatR. rubra is a C₄ plant. A further anatomical survey revealed seven otherRhynchospora species presumably having the C₄ photosynthetic pathway. In the family Cypraceae C₄ plants therefore occur in the tribe Rhynchosporeae as well as in the Scirpeae and Cypereae. The C₄ species ofRhynchospora have a normal Kranz type of leaf anatomy, although the C₄ species ofCyperus andFimbristylis presently known have an abnormal one in which the mestome sheath without chloroplasts is interposed between the Kranz tissue and the rest of the chlorenchyma. Thus inRhynchospora the Kranz tissue is in direct contact with the rest of the chlorenchyma, and it is suggested that the Kranz tissue may be homologous with the mestome sheath.     

Photosynthetic enzyme accumulation during leaf development of Arundinella hirta, a C4 grass having Kranz cells not associated with veins

The leaf of the NADP-malic enzyme type C(4) grass, Arundinella hirta, has not only mesophyll cells (MCs) and bundle sheath cells (BSCs, usual Kranz cells) but also another type of Kranz cells (distinctive cells; DCs) that are not associated with vascular bundles. We investigated photosynthetic enzyme accumulation along the base-to-tip maturation gradient of developing leaves by immunogold electron microscopy. In mature leaves, phosphoenolpyruvate carboxylase (PEPC) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) were detected in the MC cytosol and in the BSC and DC chloroplasts, respectively. Pyruvate, P(i) dikinase (PPDK) was present in the chloroplasts of all photosynthetic cells but with higher levels in the MCs. Rubisco was first detected in the basal region of emerging leaf blades where the BSCs and DCs became discernable. Subsequently, the accumulation of PEPC and PPDK was initiated in the region where the granal proliferation in the chloroplasts was conspicuous; and, suberized lamellae were formed in the cell walls of the Kranz cells. There was no difference in the patterns of cellular development and enzyme accumulation between the BSCs and DCs or between the MCs adjacent to each type of Kranz cells. These results demonstrate that, although the DCs are not associated with veins, they behaved like BSCs with respect to enzyme induction and cellular differentiation.     

Photosynthetic Characteristics of Portulaca grandiflora, a Succulent C(4) Dicot : CELLULAR COMPARTMENTATION OF ENZYMES AND ACID METABOLISM

The succulent, cylindrical leaves of the C₄ dicot Portulaca grandiflora possess three distinct green cell types: bundle sheath cells (BSC) in radial arrangement around the vascular bundles; mesophyll cells (MC) in an outer layer adjacent to the BSC; and water storage cells (WSC) in the leaf center. Unlike typical Kranz leaf anatomy, the MC do not surround the bundle sheath tissue but occur only in the area between the bundle sheath and the epidermis. Intercellular localization of photosynthetic enzymes was characterized using protoplasts isolated enzymatically from all three green cell types.