Origins of domestication and polyploidy in oca (Oxalis tuberosa : Oxalidaceae): nrDNA ITS data

As part of a study aimed at elucidating the origins of the octoploid tuber crop "oca," Oxalis tuberosa, DNA sequences of the internal trancribed spacer of nuclear ribosomal DNA (nrDNA ITS) were determined for oca and several wild Oxalis species, mostly from Bolivia. Phylogenetic analysis of these data supports a group of these species as being close relatives of oca, in agreement with morphology and cytology, but at odds with traditional infrageneric taxonomy. Variation in ITS sequences within this group is quite low (0-7 substitutions in the entire ITS region), contrasting with the highly divergent (unalignable in some cases) sequences within the genus overall. Some groups of morphologically differentiated species were found to have identical sequences, notably a group that includes oca, wild populations of Oxalis that bear small tubers, and several other clearly distinct species. The presence of a second, minor sequence type in at least some oca accessions suggests a possible contribution from a second genome donor, also from within this same species group. ITS data lack sufficient variation to elucidate the origins of oca precisely, but have identified a pool of candidate species and so can be used as a tool to screen yet unsampled species for possible progenitors.     

The Breeding System of Oxalis squamata,a Tristylous South American Species

Oxalis squamata is a perennial endemic herb of the Chilean-Argentine Andes previously reported as a distylous species. In this paper, analysis of floral morphology, incompatibility relations and population structure show that O. squamata possesses morphological and functional tristyly. The three floral morphs differ in calyx and corolla length, style and stamen length, pollen grain size and stylar pubescence. Results of hand pollinations showed the presence of a strong incompatibility system. Legitimate pollinations with the L and M morphs as seed parents produced approximately equal numbers of seeds per pollination. The S morph produced less than half the number of seeds per legitimate pollination compared to the other two morphs. Intra- and intermorph illegitimate pollinations produce almost no seeds. Morph representation in the population was isoplethic. Fruit and seed production did not differ significantly for the three morphs in the population. Comparison of experimental and natural production of fruits and seeds suggests that seed set in the natural population may be pollen-limited for the L and M morphs.     

Congruence between morphological and molecular markers inferred from the analysis of the intra-morphotype genetic diversity and the spatial structure of Oxalis tuberosa Mol.

Oxalis tuberosa is an important crop cultivated in the highest Andean zones. A germplasm collection is maintained ex situ by CIP, which has developed a morphological markers system to classify the accessions into morphotypes, i.e. groups of morphologically identical accessions. However, their genetic uniformity is currently unknown. The ISSR technique was used in two experiments to determine the relationships between both morphological and molecular markers systems. The intra-morphotype genetic diversity, the spatial structures of the diversity and the congruence between both markers systems were determined. In the first experience, 44 accessions representing five morphotypes, clearly distinct from each other, were analyzed. At the molecular level, the accessions exactly clustered according to their morphotypes. However, a genetic variability was observed inside each morphotype. In the second experiment, 34 accessions gradually differing from each other on morphological base were analyzed. The morphological clustering showed no geographical structure. On the opposite, the molecular analysis showed that the genetic structure was slightly related to the collection site. The correlation between both markers systems was weak but significant. The lack of perfect congruence between morphological and molecular data suggests that the morphological system may be useful for the morphotypes management but is not appropriate to study the genetic structure of the oca. The spatial structure of the genetic diversity can be related to the evolution of the species and the discordance between the morphological and molecular structures may result from similar selection pressures at different places leading to similar forms with a different genetic background.     

3种植物花螺原体的分离及其基本特性

[目的]调查我国植物花上的螺原体的存在,搜集我国的螺原体资源,并研究它们的基本生物学特性.[方法]常规螺原体分离、培养方法,应用暗视野显微镜和透射电子显微镜观察螺原体形态,根据16S rDNA和ITS序列构建系统发育树研究螺原体分离菌株可能的分类地位.[结果]分别从油菜(Brassica napus)、杜鹃(Rhododendron simsii)、红花酢浆草(Oxalis corymbosa)3种植物花表分离到4株螺原体CNR-1和CNR-2、CNA-1、CRW-1,对其形态、部分生理生化特性及分子生物学特性进行了初步研究.这4株螺原体在R-2液体培养基中生长良好,都能通过孔径为0.22 μm的微孔滤膜;在R-2固体培养基上呈圆形或颗粒状菌落,菌落直径约50～600 μm;在生长的某个阶段可呈典型的螺旋状,菌体直径为37.04～370.40 nm,长度约0.89～11.88 μm;它们都能利用葡萄糖作为碳源,不能利用尿素;在不含胎牛血清的R-2培养基中,它们都不能生长;菌株CNR-1、CNA-1能强烈代谢精氨酸,而CNR-2和CRW-1不能代谢精氨酸;在氨苄青霉素钠浓度高达2000 U/mL的R-2培养基中,分离菌株生长良好.根据16S rDNA序列构建的系统发育树显示,分离菌株CNR-1和CNR-2、CNA-1与蜜蜂螺原体Spiroplasma melliferum聚类较近,而CRW-1与S.clarkii聚类较近;根据ITS序列构建的系统发育树显示,CRW-1形成一个单独的分枝,其它3个菌株仍与S.melliferum聚类.[结论]以上结果初步表明,分离菌株CNR-1和CNR-2、CNA-1极有可能是spiroplasma melliferum,而CRW-1可能是一个新的螺原体种,但还需要血清学试验进一步验证.     

Chloroplast-Expressed Glutamine Synthetase (ncpGS): Potential Utility for Phylogenetic Studies with an Example from Oxalis (Oxalidaceae)

Chloroplast-expressed glutamine synthetase (ncpGS), a nuclear-encoded gene containing several introns, is introduced as a tool for phylogenetic studies at lower taxonomic levels. This gene is a member of a multigene family, but it diverged long ago from the cytosolic-expressed members of the family and appears to be single copy in the majority of taxa examined to date. The conservation of both coding sequence and position of introns has allowed the design of primers for use in a broad range of dicot taxa to amplify and sequence a region of ncpGS that contains four introns. The utility of this region in phylogenetic studies of congeneric species is illustrated by an example using eight Oxalis species. The four introns in these taxa are typical in size (76 to 136 bp), base composition (high T content), and structure (e.g., sequence of splice sites and putative branch points) for plant internal introns. Levels of variation among these ncpGS sequences compare favorably with those of the internal transcribed spacer of nuclear ribosomal DNA (ITS) from the same taxa, and results of phylogenetic analysis of ncpGS data are generally congruent with previous results using ITS.     

Oxalis jiayinensis,a new species of the Late Cretaceous from Heilongjiang,NE China

Oxalis jiayinensis, sp. nov., is described from the Late Cretaceous (TuronianSenonian) in Jiayin County, Heilongjiang Province, northeastern China. The seed is about 1 mm long, less than 1 mm in diameter, crescentic with obtuse apices and 9～10 transverse ribs. The seed wall is composed of equiaxial, strongly thickened sclereids. The fossil repre-sents the oldest occurrence of the family Oxalidaceae in the world.     

12种酢浆草属植物核型分析

【目的】为了从细胞学角度明确12种酢浆草属（Oxalis）植物核型特征与亲缘关系,探究酢浆草属植物染色体的多样性,进一步为酢浆草种质资源鉴定与杂交育种亲本选择提供参考。【方法】应用根尖压片法对酢浆草属12个物种植物有丝分裂中期的染色体形态、数目及其核型进行观察分析。【结果】结果表明：（1）7个物种的染色体数目为首次报道,5种已报道物种中,兔耳酢浆草（O. fabaefolia）和黄花酢浆草（O. pes-caprae）染色体数目与前人报道一致,其余3种,与先前报道存在差异,其中2n=26为酢浆草属植物中首次报道的染色体数目,共发现7种染色体基数,其中x=13为首次报道,倍性范围从2x~6x;实际染色体大小范围为0.27~2.23 μm,着丝点位置为中部着丝点区（m）和亚中部着丝点区（sm）,核型类型共4种,核型不对称系数范围为56.31%~65.40%。（2）12种酢浆草属植物中兔耳酢浆草最为进化,扁平酢浆草（O. compressa）最为原始。（3）根据染色体核型相似性可将12个酢浆草属植物划分为4组。【结论】研究认为,12个酢浆草属物种具有广泛的核型多样性,核型分类结果与前人形态学分类不完全一致。     

Fluctuations in heteromorphic self-incompatibility systems

Summary The genetic determination of heterostyly is briefly reviewed and experimental data are presented on the inheritance of tristyly in Oxalis compressa. It is shown that the model appropriate to Lythrum salicaria, of two diallelic loci, is inadequate to explain segregation patterns found in O. compressa, especially the reversal of dominance of the short phenotype. An extended model having an additional allele at the short locus, and a separate modifier gene, is presented and its deterministic genotype frequency dynamics examined numerically. It is shown that fixation of these extra genes or alleles is unlikely. Problems of testing the extended model are considered.     

Suitability of AFLP markers for the study of genomic relationships within theOxalis tuberosa alliance

O. tuberosa is an Andean crop that belongs to the worldwide distributed genusOxalis. On the basis of their chromosome numbers the following species (O. herrerae, O. lotoides, O. medicaginea, O. mollissima, O. oblongiformis, O. peduncularis, O. spiralis, O. subintegra, O. tabaconanensis, O. tuberosa, O. villosula) were placed in an alliance. To analyse five species belonging to theOxalis tuberosa alliance (O. oblongiformis, O. peduncularis, O. tabaconanensis, O. tuberosa andO. villosula) and a distant member of the genus (O. articulata), we examined 253 AFLP markers generated after amplification using four primer combinations. Within the alliance, two main clusters were observed, one containing the diploid species and the other group with the polyploid speciesO. tuberosa. All of the primer combinations assayed showed the same clustering pattern. Grouping of accessions of each species by data analysis corresponded largely with their previous taxonomic classifications. The concordance between the clustering of the individuals belonging to different species obtained in this work show the appropriateness of AFLP markers for this type of study. The results obtained are in good agreement with the cytogenetic hypothesis and showed a clustering behaviour, which is similar to the one previously obtained using ITS rDNA nucleotide sequence comparison.     

Stigma and style anatomy and ultrastructure in Italian Oxalis pes-caprae L. and their possible connection with self-incompatibility

The South-African Oxalis pes-caprae with trimorphic flowers is naturalised in many Mediterranean countries. In Italy only short-styled (S) populations are known for certain and the plant is believed to reproduce only asexually, due to self- and intramorph-incompatibility. This study aims to clarify anatomical, biochemical and ultrastructural features of the stigma and style of an Italian S population, also to define their possible role in incompatibility. Additional observations were also carried out on other Italian population and on short-, long- (L) and mid-styled (M) flowers from plants of South African origin. Morphological and biochemical features of flowers collected in different phenological stages during the whole flowering season were observed both under LM and TEM. In S flowers, three different zones could be distinguished in each stigma-style complex: zone I (stigmatic), zone II (substigmatic), zone III (stylar). The main differences concern the transmitting tissue: in zones I and III this is composed of loosely arranged cells with thick walls, with an abundant soft matrix which at anthesis is rich in mucopolysaccharides and lacks pectins. In zone II, it is more compact, with a less abundant wall matrix, at anthesis containing both mucopolysaccharides and pectins. In S flowers, subjected to illegitimate pollination, many pollen tubes penetrate the stigmatic papillae but apparently are arrested in zone II; only few—and mostly at the end of flowering period—succeed reaching zone III, where they encounter no further resistance to growth. Differently, after legitimate pollinations, pollen tubes succeed in crossing transmitting tissue of zone II, where cell walls of cells lying close to pollen tubes show a considerably reduced pectin content. In L and M flowers of South African origin, no peculiar transmitting tissue could be noticed in the substigmatic zone. In such flowers, pollen tubes seemingly grow easily from stigma to style both after legitimate and illegitimate pollinations. Results suggest that in S flowers the rigid transmitting tissue in zone II acts as a mechanical barrier for illegitimate pollen tubes, as the spaces between cells are narrower than the tube diameter and pectins maintain the rigidity of the cell walls, preventing cells from separating from one another. This obstacle can be overcome by legitimate pollen tubes, which make their way between cells, possibly releasing or activating specific pectinases. However, the blocking of illegitimate tubes is not absolute: a few of such tubes grow beyond zone II and reach the ovules, so that occasional fertilisation and embryo formation can be observed. In M and L flowers, different self-incompatibility mechanisms can be hypothesised.