Setaria viridis and Setaria italica, model genetic systems for the Panicoid grasses

Setaria italica and its wild ancestor Setaria viridis are diploid C(4) grasses with small genomes of ～515 Mb. Both species have attributes that make them attractive as model systems. Setaria italica is a grain crop widely grown in Northern China and India that is closely related to the major food and feed crops maize and sorghum. A large collection of S. italica accessions are available and thus opportunities exist for association mapping and allele mining for novel variants that will have direct application in agriculture. Setaria viridis is the weedy relative of S. italica with many attributes suitable for genetic analyses including a small stature, rapid life cycle, and prolific seed production. Setaria sp. are morphologically similar to most of the Panicoideae grasses, including major biofuel feedstocks, switchgrass (Panicum virgatum) and Miscanthus (Miscanthus giganteus). They are broadly distributed geographically and occupy diverse ecological niches. The cross-compatibility of S. italica and S. viridis also suggests that gene flow is likely between wild and domesticated accessions. In addition to serving as excellent models for C(4) photosynthesis, these grasses provide novel opportunities to study abiotic stress tolerance and as models for bioenergy feedstocks.     

Inheritance of some Mendelian factors in intra- and interspecific crosses between Setaria italica and Setaria viridis

Summary The inheritance of seed coat color, pericarp color, polyphenoloxidase activity and bristle, glume, collar, and leaf-base anthocyanic colorations was investigated using intra- and interspecific crosses between Setaria italica and S. viridis. The results were compared to inheritance results obtained by previous authors. In most cases, the inheritance is simple (one or two loci) and data from different crosses (intra- and interspecific) and from different authors can be compared. Two sets of two characters were found to share common loci: the polyphenoloxidase locus is one of the loci responsible for seed coat color, and bristle and glume color are determined by the same two loci. The evolutionary significance of these results is discussed.     

Setaria genome sequencing: an overview

The genus Setaria includes two important C₄ Panicoid grass species, namely S. italica (cultivated) and S. viridis (weed; wild ancestor), which together represent an appropriate model system for architectural, physiological, evolutionary, and genomic studies in related grasses. It is a diploid, inbreeder, self-fertile annual cereal grass having short life cycle and minimal growth requirements. There close relatedness to biofuel crops like switch grass and napier grass further signifies their importance. Further, foxtail millet is an important food and fodder grain crop grown in arid and semi-arid regions in many parts of the world. Therefore, an increasing interest in these species has led to a gradual accumulation and development of genomic data and genetic resources. Setaria genome sequencing is an outcome of such endeavors. These sequencing efforts uncovered several distinctive attributes of Setaria genome that may help in understanding its physiology, evolution and adaptation. This will not only aid in comparative genomics studies of Setaria and related crops including bioenergy grasses but also help in rapid advancements of genomics information for developing varieties with superior traits either through marker-assisted selection (MAS) or using transgenic approaches in these crops.     

狗尾草总DNA的提取与纯化研究

狗尾草杂草中富含多糖和酚类物质,而用已报道的提取方法均难得到高纯度DNA。为此,对提取植物DNA的SDS法进行了改进。用4．5ml细胞提取液,1．0ml饱和NaAc溶液,0．10倍样液体积的无水乙醇,氯仿／异戊醇溶液与样液体积比为0．80,与样液等体积的酚／氯仿／异戊醇溶液(28：21：1)和异丙醇,在65℃水浴加热60min,可从1．0g狗尾草中提取780μg纯DNA。     

狗尾草属5种植物叶片的形态特征

为了解狗尾草属(Setaria Beauv.)植物的叶片形态特征,采用石蜡切片法和徒手切片法观察5 种狗尾草属植物的叶片形态结构。结果表明,5 种狗尾草属植物叶表皮的解剖特征较一致,叶表皮细胞长度、气孔器大小、叶厚、角质层厚度等性状在种间存在显著差异,其中长细胞形状、垂周壁深浅、气孔器副卫细胞的形状和刺毛数量可以用来区分种类。叶片性状的相关分析表明,叶片气孔密度与气孔长度以及气孔宽度呈显著负相关,泡状细胞厚度与角质层厚度以及上表皮厚度呈显著正相关。根据禾本科叶表皮特征的演化趋势,狗尾草(S. viridis)最原始,金色狗尾草(S. glauca)和大狗尾草(S. faberii)较进化,皱叶狗尾草(S.plicata)和莩草(S. chondrachne)最高级。这些为狗尾草属植物分类学和系统演化关系提供资料。     

Phytolith analysis for differentiating between foxtail millet (Setaria italica) and green foxtail (Setaria viridis)

Foxtail millet (Setaria italica) is one of the oldest domesticated cereal crops in Eurasia, but identifying foxtail millets, especially in charred grains, and differentiating it from its wild ancestor, green foxtail (Setaria viridis), in the archaeobotanical remains, is still problematic. Phytolithic analysis provides a meaningful method for identifying this important crop. In this paper, the silicon structure patterns in the glumes, lemmas, and paleas from inflorescence bracts in 16 modern plants of foxtail millet and green foxtail from China and Europe are examined using light microscopy with phase-contrast and a microscopic interferometer. Our research shows that the silicon structure of ΩIII from upper lemmas and paleas in foxtail millet and green foxtail can be correspondingly divided into two groups. The size of ΩIII type phytolith of foxtail millet is bigger than that from green foxtail. Discriminant function analysis reveals that 78.4% of data on foxtail millet and 76.9% of data on green foxtail are correctly classified. This means certain morphotypes of phytoliths are relatively reliable tools for distinguishing foxtail millet from green foxtail. Our results also revealed that the husk phytolith morphologies of foxtail millets from China and Eastern Europe are markedly different from those from Western Europe. Our research gives a meaningful method of separating foxtail millet and green foxtail. The implications of these findings for understanding the history of foxtail millet domestication and cultivation in ancient civilizations are significant.     

UeberPanicum ambiguum Guss (sub Setaria)

Ohne Zusammenfassung     

Spike‐dip transformation of Setaria viridis

Traditional method of Agrobacterium‐mediated transformation through the generation of tissue culture had limited success for Setaria viridis, an emerging C4 monocot model. Here we present an efficient in planta method for Agrobacterium‐mediated genetic transformation of S. viridis using spike dip. Pre‐anthesis developing spikes were dipped into a solution of Agrobacterium tumefaciens strain AGL1 harboring the β‐glucuronidase (GUS) reporter gene driven by the cauliflower mosaic virus 35S (CaMV35S) promoter to standardize and optimize conditions for transient as well as stable transformations. A transformation efficiency of 0.8 ± 0.1% was obtained after dipping of 5‐day‐old S3 spikes for 20 min in Agrobacterium cultures containing S. viridis spike‐dip medium supplemented with 0.025% Silwet L‐77 and 200 μm acetosyringone. Reproducibility of this method was demonstrated by generating stable transgenic lines expressing β‐glucuronidase plus (GUSplus), green fluorescent protein (GFP) and Discosoma sp. red fluorescent protein (DsRed) reporter genes driven by either CaMV35S or intron‐interrupted maize ubiquitin (Ubi) promoters from three S. viridis genotypes. Expression of these reporter genes in transient assays as well as in T1 stable transformed plants was monitored using histochemical, fluorometric GUS activity and fluorescence microscopy. Molecular analysis of transgenic lines revealed stable integration of transgenes into the genome, and inherited transgenes expressed in the subsequent generations. This approach provides opportunities for the high‐throughput transformation and potentially facilitates translational research in a monocot model plant.     

狗尾草属野生近缘种的染色体鉴定

对来自多个国家和地区的10个种(青狗尾草S.viridis,法氏狗尾草S.faberii,轮生狗尾草S.verticillata、S.verticillifor-mis,金色狗尾草S.glauca、S.pumila、S.grisebachii、S.leucopila、S.parviflora、S.queenslandica等)50份狗尾草材料进行了染色体计数及倍性鉴定。发现狗尾草属中的青狗尾草均为二倍体,金色狗尾草S.glauca有四倍体和八倍体,轮生狗尾草S.verticillata有二倍体和四倍体,法氏狗尾草S.faberii为四倍体,S.pumila有二倍体和四倍体,S.grisebachii为二倍体,S.leucopila为二倍体,S.queenslandica为四倍体。本研究中对S.grisebachii、S.leucopila、S.queenslandica3个种是首次染色体倍性观察。发掘近缘种的有益基因是作物育种的重要途径之一,本研究搜集的谷子近缘野生种对谷子远缘杂交育种和谷子起源进化分析有重要意义。     

Histochemical enzyme pattern of microfilaria of Setaria cervi

The present study deals with the histochemical localization of glucose-6-phosphatase, malic dehydrogenase and aldolase in the microfilaria of Setaria cervi. Marked activity of glucose-6-phosphatase was observed in the cephalic cells, excretory and anal pores, G-cells and Innenk?rper. Malic dehydrogenase activity was noted throughout the body (including cuticle) of the microfilaria except for Innenk?rper. Intense aldolase activity was observed in the excretory pore and G-cells only. Muscle cells and anal pore were negative for this enzyme.     

Cuticle specific antigens from filarial parasite Setaria digitata

A fairly clean antigenic cuticle was isolated from the S. digitata by dissection. Crossed immunoelectrophoresis of cuticular antigens against rabbit antiserum to cuticular antigens gave 30 anodic and 5 cathodic precipitin arcs. The cuticle antiserum cross reacted with muscle, uterus and pseudocoelomic fluid. When the antiserum was absorbed individually with these cross reacting somatic preparations, analysis against cuticle antigens gave only a limited number of precipitin arcs. But the results are clear enough to indicate the presence of cuticle specific antigens. As the cuticle of the parasite is in contact with the host system, an antigenic preparation from it may prove a useful tool for the detection of filariasis.     

Reference genome sequence of the model plant Setaria

We generated a high-quality reference genome sequence for foxtail millet (Setaria italica). The ～400-Mb assembly covers ～80% of the genome and >95% of the gene space. The assembly was anchored to a 992-locus genetic map and was annotated by comparison with >1.3 million expressed sequence tag reads. We produced more than 580 million RNA-Seq reads to facilitate expression analyses. We also sequenced Setaria viridis, the ancestral wild relative of S. italica, and identified regions of differential single-nucleotide polymorphism density, distribution of transposable elements, small RNA content, chromosomal rearrangement and segregation distortion. The genus Setaria includes natural and cultivated species that demonstrate a wide capacity for adaptation. The genetic basis of this adaptation was investigated by comparing five sequenced grass genomes. We also used the diploid Setaria genome to evaluate the ongoing genome assembly of a related polyploid, switchgrass (Panicum virgatum).     

一种狗尾草病原真菌的鉴定及菌株致病性研究

经形态学鉴定和rDNA ITS序列分析,16株分离自北京、河北、河南发病狗尾草的菌株、2株分别分离自河南发病虎尾草、牛筋草的菌株和1株分离自青海发病野燕麦的菌株被鉴定为狗尾草平脐蠕孢Bipolaris setariae。接种试验表明,来自狗尾草的菌株比来自其他寄主植物的菌株对狗尾草致病性强,分离自野燕麦的菌株对狗尾草无致病性,分离自不同地区不同样品狗尾草的菌株其致病性有显著差异。菌株NY1对狗尾草有很强致病性,接种后5d植株叶片即全部呈枯死状,接种后7d整个植株枯萎死亡。菌株NY1对马唐和虎尾草也有很强致病性,但对于大多数供试栽培植物致病性很弱或无致病性。因此,B. setariae NY1菌株具有进一步开发成为狗尾草、马唐和虎尾草等杂草的生物除草剂的潜力。     

Excretory secretory antigens of filarial parasite Setaria digitata

Excretory Secretory (ES) material isolated from the culture fluid of S. digitata was highly antigenic. Neither oesophagus nor excretory cells and excretory pore of the parasite showed reasonable fluorescence with ES antisera. However, the uterine tissue and the egg showed strong fluorescence. The egg showed fluorescence mainly in the space between embryo and egg membrane (amniotic fluid). The amniotic fluid was highly antigenic and appears to be the most important source of ES material released by the filarial parasites.     

Isolation of gametes and zygotes from Setaria viridis

Setaria viridis, the wild ancestor of foxtail millet (Setaria italica), is an effective model plant for larger C₄ crops because S. viridis has several desirable traits, such as short generation time, prolific seed production and a small genome size. These advantages are well suited for investigating molecular mechanisms in angiosperms, especially C₄ crop species. Here, we report a procedure for isolating gametes and zygotes from S. viridis flowers. To isolate egg cells, ovaries were harvested from unpollinated mature flowers and cut transversely, which allowed direct access to the embryo sac. Thereafter, an egg cell was released from the cut end of the basal portion of the dissected ovary. To isolate sperm cells, pollen grains released from anthers were immersed in a mannitol solution, resulting in pollen-grain bursting, which released sperm cells. Additionally, S. viridis zygotes were successfully isolated from freshly pollinated flowers. Isolated zygotes cultured in a liquid medium developed into globular-like embryos and cell masses. Thus, isolated S. viridis gametes, zygotes and embryos are attainable for detailed observations and investigations of fertilization and developmental events in angiosperms.