Phylogenetic analysis of tribe Salsoleae (Chenopodiaceae) based on ribosomal ITS sequences: implications for the evolution of photosynthesis types.

Diversity in photosynthetic pathways in the angiosperm family Chenopodiaceae is expressed in both biochemical and anatomical characters. To understand the evolution of photosynthetic diversity, we reconstructed the phylogeny of representative species of tribe Salsoleae of subfamily Salsoloideae, a group that exhibits in microcosm the patterns of photosynthetic variation present in the family as a whole, and examined the distribution of photosynthetic characters on the resulting phylogenetic tree. Phylogenetic relationships were inferred from parsimony analysis of nucleotide sequences of the internal transcribed spacer regions (ITS) of the 18S-26S nuclear ribosomal DNA of 34 species of Salsola and related genera (Halothamnus, Climacoptera, Girgensohnia, Halocharis, and Haloxylon) and representative outgroups from tribes Camphorosmeae (Camphorosma lessingii, Kochia prostrata, and K. scoparia) and Atripliceae (Atriplex spongiosa). A highly resolved strict consensus tree largely agrees with photosynthetic type and anatomy of leaves and cotyledons. The sequence data provide strong support for the origin and evolution of two main lineages of plants in tribe Salsoleae, with NAD-ME and NADP-ME C(4) photosynthesis, respectively. These groups have different C(4) photosynthetic types in leaves and different structural and photosynthetic characteristics in cotyledons. Phylogenetic relationships inferred from ITS sequences generally agree with classifications based on morphological data, but deviations from the existing taxonomy were also observed. The NAD-ME C(4) lineage contains species classified in sections Caroxylon, Malpigipila, Cardiandra, Belanthera, and Coccosalsola, and the NADP-ME lineage comprises species from sections Coccosalsola and Salsola. Reconstruction of photosynthetic characters on the ITS phylogeny indicates separate NAD-ME and NADP-ME lineages and suggests two reversions to C(3) photosynthesis. Reconstruction of geographic distributions suggests Salsoleae originated and diversified in central Asia and subsequently dispersed to Africa, Europe, and Mongolia. Inferred patterns and processes of photosynthetic evolution in Salsoleae should further our understanding of biochemical and anatomical evolution in Chenopodiaceae as a whole.

Compartmentation of photosynthesis in cells and tissues of C(4) plants.

Critical to defining photosynthesis in C(4) plants is understanding the intercellular and intracellular compartmentation of enzymes between mesophyll and bundle sheath cells in the leaf. This includes enzymes of the C(4) cycle (including three subtypes), the C(3) pathway and photorespiration. The current state of knowledge of this compartmentation is a consequence of the development and application of different techniques over the past three decades. Initial studies led to some alternative hypotheses on the mechanism of C(4) photosynthesis, and some controversy over the compartmentation of enzymes. The development of methods for separating mesophyll and bundle sheath cells provided convincing evidence on intercellular compartmentation of the key components of the C(4) pathway. Studies on the intracellular compartmentation of enzymes between organelles and the cytosol were facilitated by the isolation of mesophyll and bundle sheath protoplasts, which can be fractionated gently while maintaining organelle integrity. Now, the ability to determine localization of photosynthetic enzymes conclusively, through in situ immunolocalization by confocal light microscopy and transmission electron microscopy, is providing further insight into the mechanism of C(4) photosynthesis and its evolution. Currently, immunological, ultrastructural and cytochemical studies are revealing relationships between anatomical arrangements and photosynthetic mechanisms which are probably related to environmental factors associated with evolution of these plants. This includes interesting variations in the C(4) syndrome in leaves and cotyledons of species in the tribe Salsoleae of the family Chenopodiaceae, in relation to evolution and ecology. Thus, analysis of structure-function relationships using modern techniques is a very powerful approach to understanding evolution and regulation of the photosynthetic carbon reduction mechanisms.

C4 plants in the vegetation of Mongolia: their natural occurrence and geographical distribution in relation to climate.

The natural geographical occurrence, carbon assimilation, and structural and biochemical diversity of species with C4 photosynthesis in the vegetation of Mongolia was studied. The Mongolian flora was screened for C4 plants by using 13C/12C isotope fractionation, determining the early products of 14CO2 fixation, microscopy of leaf mesophyll cell anatomy, and from reported literature data. Eighty C4 species were found among eight families: Amaranthaceae, Chenopodiaceae, Euphorbiaceae, Molluginaceae, Poaceae, Polygonaceae, Portulacaceae and Zygophyllaceae. Most of the C4 species were in three families: Chenopodiceae (41 species), Poaceae (25 species) and Polygonaceae, genus Calligonum (6 species). Some new C4 species in Chenopodiaceae, Poaceae and Polygonaceae were detected. C4 Chenopodiaceae species make up 45% of the total chenopods and are very important ecologically in saline areas and in cold arid deserts. C4 grasses make up about 10% of the total Poaceae species and these species naturally concentrate in steppe zones. Naturalized grasses with Kranz anatomy,of genera such as Setaria, Echinochloa, Eragrostis, Panicum and Chloris, were found in almost all the botanical-geographical regions of Mongolia, where they commonly occur in annually disturbed areas and desert oases. We analyzed the relationships between the occurrence of C4 plants in 16 natural botanical-geographical regions of Mongolia and their major climatic influences. The proportion of C4 species increases with decreasing geographical latitude and along the north-to-south temperature gradient; however grasses and chenopods differ in their responses to climate. The abundance of Chenopodiaceae species was closely correlated with aridity, but the distribution of the C4 grasses was more dependent on temperature. Also, we found a unique distribution of different C4 Chenopodiaceae structural and biochemical subtypes along the aridity gradient. NADP-malic enzyme (NADP-ME) tree-like species with a salsoloid type of Kranz anatomy, such as Haloxylon ammodendron and Iljinia regelii, plus shrubby Salsola and Anabasis species, were the plants most resistant to ecological stress and conditions in highly arid Gobian deserts with less than 100 mm of annual precipitation. Most of the annual C4 chenopod species were halophytes, succulent, and occurred in saline and arid environments in steppe and desert regions. The relative abundance of C3 succulent chenopod species also increased along the aridity gradient. Native C4 grasses were mainly annual and perennial species from the Cynodonteae tribe with NAD-ME and PEP-carboxykinase (PEP-CK) photosynthetic types. They occurred across much of Mongolia, but were most common in steppe zones where they are often dominant in grazing ecosystems.

Occurrence of C(3) and C(4) photosynthesis in cotyledons and leaves of Salsola species (Chenopodiaceae).

Most species of the genus Salsola (Chenopodiaceae) that have been examined exhibit C(4) photosynthesis in leaves. Four Salsola species from Central Asia were investigated in this study to determine the structural and functional relationships in photosynthesis of cotyledons compared to leaves, using anatomical (Kranz versus non-Kranz anatomy, chloroplast ultrastructure) and biochemical (activities of photosynthetic enzymes of the C(3) and C(4) pathways, (14)C labeling of primary photosynthesis products and (13)C/(12)C carbon isotope fractionation) criteria. The species included S. paulsenii from section Salsola, S. richteri from section Coccosalsola, S. laricina from section Caroxylon, and S. gemmascens from section Malpigipila. The results show that all four species have a C(4) type of photosynthesis in leaves with a Salsoloid type Kranz anatomy, whereas both C(3) and C(4) types of photosynthesis were found in cotyledons. S. paulsenii and S. richteri have NADP- (NADP-ME) C(4) type biochemistry with Salsoloid Kranz anatomy in both leaves and cotyledons. In S. laricina, both cotyledons and leaves have NAD-malic enzyme (NAD-ME) C(4) type photosynthesis; however, while the leaves have Salsoloid type Kranz anatomy, cotyledons have Atriplicoid type Kranz anatomy. In S. gemmascens, cotyledons exhibit C(3) type photosynthesis, while leaves perform NAD-ME type photosynthesis. Since the four species studied belong to different Salsola sections, this suggests that differences in photosynthetic types of leaves and cotyledons may be used as a basis or studies of the origin and evolution of C(4) photosynthesis in the family Chenopodiaceae.