高山榕染色体制片优化及核型分析

以高山榕种子的根尖为材料进行染色体常规压片,比较不同预处理方法和解离方法对高山榕染色体制片的影响,以选择最优的压片方法制片并进行核型分析。结果显示,预处理24 h的总体效果优于12 h;3种预处理液的总体作用效果为0.05%秋水仙素>混合液>0.002 mol.L-18-羟基喹啉,综合比较后认为混合液低温4℃处理24 h为最佳预处理方法。解离方法选择1 mol.L-1HCl中60℃水浴2~3 min较为适宜。首次报道了高山榕核型公式为2n=2x=30=22m(2SAT)+6sm+2T,核型不对称系数为61.54%,核型属于Stebbins核型分类中的2C类型。     

古生代有孔虫实体化石研究方法

古生代有孔虫大部分是通过切片进行研究的,但是普通的切片法很难得到非常好的定向标本,这就产生了很多同物异名,限制了有孔虫在古生代生物地层学、古生态学和古海洋学中的应用。另外由于口孔的内部和外部特征在分类学中的地位越来越重要,所以通过实体化石来研究古生代有孔虫理应受到更多的重视。本文论证了在古生代地层中获得实体化石的可能性,对化石提取的实验步骤进行了系统总结,细述了对实体化石进行扫描电镜和切片研究的全过程,并根据笔者的经验,对一些技术细节及注意事项进行了详细介绍。本文最后建议,古生代的有孔虫研究应该尽可能地利用实体化石来进行。     

华中五味子染色体制片优化及核型分析

比较不同预处理和解离方法对华中五味子染色体制片的影响,采用华中五味子的萌发芽、新枝茎尖、幼叶等不同部位进行染色体制片,观察500个细胞,比较不同部位中期细胞和适宜核型分析的中期细胞所占比例,结果显示,0.1%秋水仙素预处理1 h、1 mol/L盐酸常温解离12 min制片所得染色体分散效果最佳。5~15 mm幼叶侧边组织制片最适宜华中五味子核型分析。核型公式为2n=2x=28=26 m 2sm,核型不对称系数(As.k)为56.30%,属于1B类型,核型对称性程度高,表明华中五味子在进化中处于比较原始类型。     

黄果龙葵和龙葵染色体制片优化及核型分析

该实验以黄果龙葵和龙葵的根尖为实验材料,进行不同的预处理、固定和解离,确定出各种材料适合于核型分析的制片方法。结果表明:龙葵于15℃条件下经0.05%秋水仙素预处理2.5h,固定后用1mol/L HCl酸解后,染色观察,得到的染色体分散,易于染色体计数和形态观察。用此方法对黄果龙葵和龙葵进行核型分析,结果发现:黄果龙葵和龙葵都属于小型染色体,黄果龙葵为四倍体,核型公式为K(4n)=48=4sm+44m,核型不对称系数为56.22%,属于2B核型。龙葵为六倍体,核型公式为K(6n)=72=72m,核型不对称系数为55.89%,属于1B核型。     

Taxonomic significance of spine morphology in the echinoid genera Diadema and Echinothrix

Abstract. The spine morphology of all established species of Diadema and Echinothrix, including 2 color morphs of E. calamaris, were examined externally and internally via transverse sectioning to identify diagnostic species features and to assess the morphological relationship between species. Forty‐nine different morphological characters were measured and analysed using ordination by multi‐dimensional scaling (MDS) and cluster analysis. Specimens of Diadema paucispinum and D. setosum had very distinct spine structures. In D. paucispinum, the spines were more robust than those of other species of Diadema. This was evident in the spine's internal structure, with large, closely packed solid wedges, a small axial cavity, and rings of trabeculae throughout the spine's length. The spines in D. setosum were distinctive because of their length in relation to test size and the reduced flaring of their verticillations. The spines of other members of this genus were very similar to each other. Without careful sectioning, the spines from specimens of D. antillarum, D. ascensionis, D. mexicanum and D. savignyi were difficult to differentiate. The internal structures of spines for each species did, however, possess a combination of features that differentiated the species. Such features included the shape, orientation, and number of solid wedges, the presence or absence of spokes and rings of trabeculae between the solid wedges, and the presence or absence of tissue within the axial cavity. Individuals of Diadema palmeri also had spines morphologically similar to other species, however, the red pigmentation of these spines (in life and when preserved) made them easily distinguishable. The spine structures of the 2 species of Echinothrix were starkly different, while the white and brown color morphs of E. calamaris had morphologically distinctive ambulacral and interambulacral spines. The blunt, open‐tipped interambulacral spines, with reticular tissue present in the axial cavity of the white color morph, were easily distinguished from the pointed, closed‐tipped spines, with a hollow axial cavity found in the brown color morph. Such differences indicate that the brown color morph is either a subspecies or a separate species. Taken together the data show that each species has significant morphological differences in the structure of the spines. It is evident from our data that spine morphology is a useful tool to differentiate these commonly confused species.     

A lamellated striated layer in the spore wall of the smut fungus Entorrhiza (Ustilaginales)

Summary The spore wall in the smut fungus Entorrhiza has been investigated, with particular reference to layer 3. The wall is stratified into four layers, numbered 1–4 from the outside to inside of the wall. Layer 3 has a lamellated or striated organization, depending on the type of specimen preparation used for examination. In this study, thin sectioning and freeze-etching methods were used in transmission electron microscopy. Layer 3 is approximately 50 nm thick and is the narrowest layer of the wall. Thin sections viewed at high magnification show a lamellated organization, consisting of alternate electron dense and translucent spaces. Usually between 16–20 lamellae form the layer, with a lamella having a thickness of about 1–2 nm. At high resolution, each electron dense lamella is resolved as a row of closely packed subunits, approximately 1.5 nm in diameter. The electron translucent lamellae probably represent mainly lipoidal material, which is extracted during specimen preparation. In freeze-etch preparations layer 3 is termed the striated layer. Fracturing exposes face views of the layer, which at low magnification has a wrinkled appearance. At high magnification, layer 3 has a structure consisting of an irregular mosaic of striations. The striations in each area of the mosaic are arranged parallel, and regularly spaced 11–13 nm apart. Each area of the mosaic is separated from an adjacent area by a small step; this represents where the plane of fracture changed to a different level within layer 3. Fracturing probably occurs in the lipoidal region of the layer, corresponding to the electron translucent lamellae seen in thin section. At high magnification, the striations are resolved into subunit-like particles, approximately 1–2 nm in diameter. Layer 3 is believed to form an impervious region in the spore wall. Layer 3 in Entorrhiza closely resembles the partition layer reported in spores of other Tilletiaceae. This demonstration of a common wall layer strengthens the relationship between Entorrhiza and the rest of the Tilletiaceae. Entorrhiza is the only smut that forms galls on host roots.     

Maurer's cleft organization in the cytoplasm of plasmodium falciparum-infected erythrocytes: new insights from three-dimensional reconstruction of serial ultrathin sections

Maurer's clefts are single-membrane-limited structures in the cytoplasm of erythrocytes infected with the human malarial parasite Plasmodium falciparum. The currently accepted model suggests that Maurer's clefts act as an intermediate compartment in protein transport processes from the parasite across the cytoplasm of the host cell to the erythrocyte surface, by receiving and delivering protein cargo packed in vesicles. This model is mainly based on two observations. Firstly, single-section electron micrographs have shown, within the cytoplasm of infected erythrocytes, stacks of long slender membranes in close vicinity to round membrane profiles considered to be vesicles. Secondly, proteins that are transported from the parasite to the erythrocyte surface as well as proteins facilitating the budding of vesicles have been found in association with Maurer's clefts. Verification of this model would be greatly assisted by a better understanding of the morphology, dimensions and origin of the Maurer's clefts. Here, we have generated and analyzed three-dimensional reconstructions of serial ultrathin sections covering segments of P. falciparum-infected erythrocytes of more than 1 microm thickness. Our results indicate that Maurer's clefts are heterogeneous in structure and size. We have found Maurer's clefts consisting of a single disk-shaped cisternae localized beneath the plasma membrane. In other examples, Maurer' clefts formed an extended membranous network that bridged most of the distance between the parasite and the plasma membrane of the host erythrocyte. Maurer's cleft membrane networks were composed of both branched membrane tubules and stacked disk-shaped membrane cisternae that eventually formed whorls. Maurer's clefts were visible in other cells as a loose membrane reticulum composed of scattered tubular and disk-shaped membrane profiles. We have not seen clearly discernable isolated vesicles in the analyzed erythrocyte segments suggesting that the current view of how proteins are transported within the Plasmodium-infected erythrocyte may need reconsideration.     

Inner-ear structure in Morganucodon, an early Jurassic mammal

The shapes and dimensions of the cochlear cavities from four petrosals of the genus Morganucoden obtained through sectioning and reconstruction. Morganucodon dates from the early Jurassic and represents many of the earliest known mammal specimens. Each Morganucodon petrosal fossil was embedded in Araldite and shaved with an ultramicrotome to expose the internal structure. Line drawings of the exposed cross-sections were digitized and used to produce three-dimensional reconstructions. The reconstructed Monganucodon cochlear cavities differ from extant mammalian cochleas in several respects: they are uncoiled, shorter in length, and lack the bony lamina which supports the basilar membrane. These three features ar characteristic of extant Aves and Reptilia.     

Serial Sectioning of Insects with Hard Exoskeleton by Dissolution of the Exocuticle

The method reported here was designed to produce paraffin serial sections as thin as 5 Mm of insects or other arthropods with a hard cuticle. Heads and abdomens of Apis mellifera, Eristalomyia tenax and Tenebrio molitor were fixed with Schaffer's liquid, dehydrated with 80% ethanol, 90% ethanol, two changes of 100% isopropanol (2 hr each) and 12 hr in a 1:1 mixture of paraffin (58 C melting point) at 60 C. They were molded in paraffin after 12 hr of infiltration under a partial vacuum at 60 C. Large body openings of objects were sealed with paraffin to prevent infiltration of solvents.

Thereafter, the outer paraffin was removed manually and with xylene (15 min); the cuticle was rehydrated with 100% isopropanol and 100% ethanol (15 min each). The objects were then treated with Sputofluol (Merck; a mixture of NaOH and NaCIO) until they became white or their colorless endocuticle was stainable with aniline blue WS (C.I. 42755) after rinsing in a 50% acetic acid solution (v/v). They were then dehydrated with 100% ethanol and 100% isopropanol (15 min each) and subsequently re-embedded in paraffin. They were molded, sectioned, stained and mounted as usual.     

New perspectives for wood anatomical analysis in dendrosciences: The GSL1-microtome

The variability of wood anatomical features in the rings of trees and shrubs is known to be dependent on multifaceted environmental parameters. The ability to determine anatomical variations over longer time periods as decades or centuries is a step forward in dendroecological and dendroclimatological research. In this regard micro sectioning is still one of the basic requirements but sectioning devices (microtomes) designed for wood anatomical purposes are rarely available. We present an affordable heavy duty, but light-weight microtome operated with removable blades. This portable device enables the production of large sections for dendroclimatological reconstructions as well as various types of common sections for wood anatomical studies.