The spatial distribution pattern of the brown planthopperNilaparvata lugens Stål (Homoptera: Delphacidae) in west Java,Indonesia

Spatial distribution pattern of the brown planthopper (BPH) was analyzed at 9 experimental fields in the northern part of West Java during two consecutive rice cropping seasons, i.e., wet and dry seasons. The population of each developmental stage and wing form of BPH at each location showed consistent departure from the random (Poisson) distribution, the variances of the densities in most cases exceeding their means. Namely, the distribution pattern of BPH per hill of rice plant was found to have a general tendency to be aggregated or contagious and to fit fairly well to the negative binomial model. The tendency for aggregation was further confirmed by both the β-values of -m regression being larger than unity and the C_A-values being larger than zero for each developmental stage. Although significant variations in the distribution pattern as measured by β- or C_A-value were observed between different developmental stages, between wing forms and among locations, the degree of aggregation for a given developmental stage at each experimental field remained fairly stable throughout the crop period, despite wide temporal changes in population density. Possible factors to explain these characteristics of the spatial distribution pattern of the BPH in West Java were discussed with reference to the process generating it.     

Large-Scale Temporal Changes in Spatial Pattern During Declines of Abundance and Occupancy in a Common Moth

We examined the changes in spatial pattern that accompanied the population decline of the garden tiger moth, Arctia caja (L.) in Britain between 1968 and 1999 using Spatial Analysis by Distance Indices (SADIE) techniques. A principal co-ordinate analysis of all pair-wise spatial associations between years indicated three groupings of years: 1969–1978,1979–1990,1991–1999, which revealed three phases during the population decline: an early period with highly structured spatial pattern; a middle period with nearly random distribution overall; and a recent period with highly structured spatial pattern but small abundance overall. The change in spatial structure in the early 1980’s accompanied rapid changes in abundance but preceded a sharp decline in occupancy and confirms that the sharp decline in abundance included decreases from widespread high-density sites. Perhaps unusually, A. caja varied from spatially aggregated to randomly distributed and back to spatially aggregated, all while its abundance declined sharply. Present distribution pattern may reflect past abundance changes poorly in this species. Areas showing the greatest variation in abundance displayed the greatest range in spatial structure, but also the greatest stability of spatial pattern, indicating changes between extremes of spatial pattern occurred slowly. SADIE techniques are a powerful method to quantify temporal changes in spatial pattern and relate them to temporal changes in abundance.     

Aggregation patterns ofBemisia tabaci in response to insecticide applications

We measured the effect of insecticide applications on the distribution patterns of the sweet potato whitefly,Bemisia tabaci (Gennadius), in fields of cantaloupe,Cucumis melo L., in Yuma, Arizona. Whitefly infestations were measured by counts of adults, eggs, first to mid/fourth instars, and late-fourth instars (‘red-eyed’) nymphs. Adults were sampled from the entire leaf, and immature stages were counted in a 1-cm² area of a leaf. The indicesb (Power Law), β (Patchiness regression) and I _d (Morisita) indicated that all life stages were aggregated, but results for the three indices were not similar for determining the relative aggregation levels between treatments. In general, indices β andI _d indicated higher aggregation in insecticide-treated fields, whereasb had mixed results. The Morisita index was sensitive to a few unusually high means among a series of low densities in the treatment plots, what could be attributed to refuges due to failure in the insecticide applications. Despite the usefulness of the Power Law and the Patchiness regression for describing the relationship between spatial or temporal variability and mean densities, we suggest thatI _d is more appropriate for expression of spatial distribution because it is based on a precise definition of aggregation.     

Variation in demographic and fine-scale genetic structure with population-history stage of <Emphasis Type="Italic">Hemerocallis taeanensis</Emphasis> (Liliaceae) across the landscape

Fine-scale genetic structure (FSGS) in plant populations is expected to be influenced by variation in demographic processes across space and over time. I chose Hemerocallis taeanensis (Liliaceae), a perennial herb with a rapid population turnover, to quantify how demographic structure and FSGS change with a population’s history (i.e., density). Nonaccumulative O-ring statistic and spatial autocorrelation analysis (kinship coefficient, F _ij ) were used to quantify spatial patterns of individuals and FSGS in four populations belonging to two population stages (expansion and maturation) in west-central Korea. The O-ring function revealed that significant aggregation of individuals occurs at short spatial scales during the earlier stage of population expansion, which reflects restricted seed dispersal around maternal individuals. However, this pattern disappears as the population density increases during population maturation, probably due to a high population density. Significant evidence of FSGS was found in two populations at the stage of population expansion (Sp, a statistic which describes the rate of decrease of pairwise kinship with distance, was 0.018 and 0.029). The results show that most seeds fall around maternal plants when initially established colonists proliferate at suitable microhabitats. In contrast to this, much lower Sp values (−0.003 and 0.004) were estimated for two populations at the stage of population maturation, which may result from the overlapping of seed shadows due to high adult density. All of these results demonstrate considerable variation in within-population demographic and genetic structures of H. taeanensis with respect to population temporal stage across the landscape.     

A distance measure of spatial pattern in a biological population

Suppose that n individuals locate independently and randomly on a segment of line of finite length (habitat). Let the theoretical and observed ranges of the sites of the individuals on the segment be μ_n-1 and r_n-1, respectively. Then, the degree of dispersion of the individual sites is measured by the ratio, T_n=n_n-1/μ_n-1, as follows: A random spatial pattern for I_r−1=1 An aggregated spatial pattern for 0≤I_r<1 A uniform spatial pattern for (n+1)/(n−1)≥I_r>1. Another method was derived. Let the probability that an observed range is less than r_n−1 be I_p, under the hypothesis of a Beta distribution. Then indicates A random spatial pattern for I_p=1/2 An aggregated spatial pattern for I_p<1/2 A uniform spatial pattern for I_p>1/2. The first index can be used for comparing populations having the same number of individuals, whereas the second one can be used for comparing populations with different numbers of individuals.     

濒危植物狭果秤锤树群落内主要树种的空间分布格局和关联性

为了解狭果秤锤树(Sinojackia rehderiana)群落的物种共存机制,研究了群落0.5hm~2样地内5主要树种的空间分布格局和种间关联性。结果表明,香樟(Cinnamomum camphora)、紫弹树(Celtis biondii)处于群落上层,尾叶冬青(Ilex wilsonii)在群落中层,下层以狭果秤锤树、瓜木(Alangium platanifolium)为主。狭果秤锤树、香樟、尾叶冬青、瓜木在小尺度上均为聚集分布,随着尺度的增大呈现出均匀分布或随机分布的趋势;紫弹树在全部尺度上为随机分布。尾叶冬青、瓜木在较小尺度上与狭果秤锤树的空间分布呈负相关,而香樟、紫弹树与狭果秤锤树则在较大尺度上呈负相关性,香樟、紫弹树、尾叶冬青、瓜木两两之间均无明显空间相关性。因此,推断密度制约和种子扩散限制对狭果秤锤树群落空间格局形成起作用,生境异质性的影响没有明显表现出来,群落尚未达到稳定状态。     

Overexpression of rice phytochrome A in tobacco seedlings modifies responsiveness but not competence to phytochrome

To analyse the control of rice phytochrome A (phyA) overexpression (wild type or variously mutated) on gene regulation, transgenic tobacco lines overexpressing various rice phyA constructs were crossed with transgenic tobacco lines containing mustard Lhcb1 or Chs1 promoters fused to the uidA reporter gene (-glucuronidase). It was demonstrated that the temporal pattern of competence to respond to phytochrome was not altered by rice phyA overexpression. Also, overexpression of rice phyA did not change the spatial pattern of gene expression. The responsiveness to red and far-red light, on the other hand, depended on the type of overexpressed rice phyA in a structure-function relation: the serine-to-alanine mutant mediated an enhanced response both under continuous red and far-red light, whereas the N-terminal deletion mutant showed a dominant negative effect under continuous far-red light and even after red light pulses. However, the effectiveness of rice phyA overexpression depended on the promoter construct and the developmental stage of the seedlings. The Lhcb1 promoter also conferred -glucuronidase activity in etiolated seedlings. This dark expression could be decreased by a long-wavelength farred light pulse given early in development (24 h after sowing), indicating that this phenomenon is under the control of stable types of phytochrome.Abbreviations Chs1 chalcone synthase - GUS -glucuronidase - Lhcb1 type 1 light-harvesting chlorophyll a/b-binding protein - NTD N-terminal deletion mutant of rice phyA - phyA phytochrome A - phyB phytochrome B - Pfr far-red absorbing form of phytochrome - Pr red-absorbing form of phytochrome - RW rice wild-type phyA - S/A serine-to-alanine mutant of rice phyA - XAN wild-type tobacco cv. Xanthi We thank N.-H. Chua (Rockefeller Univ., New York, USA) and J. Stockhaus (Heinrich-Heine-Universität, Düsseldorf, Germany) for providing seeds from tobacco lines overexpressing the diverse rice phyA proteins. The work was supported by a grant from the Human Frontier Science Program and a grant from Deutsche Forschungsgemeinschaft (SFB 388). K.E. is a recipient of a Landesgraduierten-förderung fellowship     

Atrial proarrhythmia due to increased inward rectifier current (IK1) arising from KCNJ2 mutation – A simulation study

Atrial fibrillation (AF) has been linked to increased inward rectifier potassium current, I_K1, either due to AF-induced electrical remodelling, or from functional changes due to the Kir2.1 V93I mutation. The aim of this simulation study was to identify at cell and tissue levels' mechanisms by which increased I_K1 facilitates and perpetuates AF. The Courtemanche et al. human atrial cell action potential (AP) model was modified to incorporate reported changes in I_K1 induced by the Kir2.1 V93I mutation in both heterozygous (Het) and homozygous (Hom) mutant forms. The modified models for wild type (WT), Het and Hom conditions were incorporated into homogeneous 1D, 2D and 3D tissue models. Restitution curves of AP duration (APD), effective refractory period (ERP) and conduction velocity (CV) were computed and both the temporal and the spatial vulnerability of atrial tissue to re-entry were measured. The lifespan and tip meandering pattern of re-entry were also characterised. For comparison, parallel simulations were performed by incorporating into the Courtmanche et al. model a linear increase in maximal I_K1 conductance. It was found that the gain-in-function of V93I ‘mutant’ I_K1 led to abbreviated atrial APs and flattened APD, ERP and CV restitution curves. It also hyperpolarised atrial resting membrane potential and slowed down intra-atrial conduction. V93I ‘mutant’ I_K1 reduced the tissue's temporal vulnerability but increased spatial vulnerability to initiate and sustain re-entry, resulting in an increased overall susceptibility of atrial tissue to arrhythmogenesis. In the 2D model, spiral waves self-terminated for WT (lifespan < 3.3 s) tissue, but persisted in Het and Hom tissues for the whole simulation period (lifespan > 10 s). The tip of the spiral wave meandered more in WT tissue than in Het and Hom tissues. Increased I_K1 due to augmented maximal conductance produced similar results to those of Het and Hom Kir2.1 V93I mutant conditions. In the 3D model the dynamic behaviour of scroll waves was stabilized by increased I_K1. In conclusion, increased I_K1 current, either by the Kir2.1 V93I mutation or by augmented maximal conductance, increases atrial susceptibility to arrhythmia by increasing the lifespan of re-entrant spiral waves and the stability of scroll waves in 3D tissue, thereby facilitating initiation and maintenance of re-entrant circuits.     

北京山区栓皮栎林优势种群点格局分析

采用点格局分析方法对北京山区栓皮栎林主要种群和不同发育阶段种群的空间分布格局及相互关系进行分析。结果表明:(1)群落物种组成简单,栓皮栎和油松是群落的优势种群,优势种群以中径木为主,缺乏可供更新的幼苗,种群呈现出衰退趋势;(2)在研究尺度内,栓皮栎、油松单种的空间分布格局以聚集型分布为主,随着尺度的增大呈现出随机分布和均匀分布的趋势;栓皮栎和油松呈现显著负相关;(3)优势种群不同发育阶段的空间分布格局与研究尺度紧密相关,栓皮栎小树和中树以聚集型分布为主、大树呈随机分布,而油松不同发育阶段均以随机分布为主;(4)栓皮栎、油松种群不同发育阶段之间的空间关联主要发生在小尺度范围内,小树和中树以正关联为主,而中树和大树、小树和大树以负关联为主;随着研究尺度的增大趋于无关联。     

桥山林区麻栎种群不同发育阶段空间格局及关联性

为探究麻栎种群不同发育阶段的空间分布特征及关联性,以黄土高原南部桥山林区麻栎天然次生林为研究对象,用L(r)函数对麻栎种群不同发育阶段空间分布特征及关联性进行分析。结果表明:(1)样地内麻栎种群各发育阶段数量结构均为不规则倒"J"型,种群结构稳定,能实现持续更新。(2)麻栎种群总体的空间分布特征受生境异质性影响较大,纯林样地中,种群在0—10 m尺度上表现为高强度聚集分布,5—35 m尺度表现为低强度聚集;混交林样地中,0—5 m尺度上表现为高强度聚集分布,5—35 m尺度上则表现为低强度聚集分布。(3)随着种群的发育及尺度的增大,麻栎种群空间聚集强度减弱,纯林样地内,麻栎幼苗在0—6 m的小尺度表现为聚集分布,6—12 m尺度上表现为随机分布;幼树在各尺度上均表现为聚集分布;中龄树与成龄树在各尺度上均表现为随机分布。混交林样地中,仅幼苗在0—6 m尺度上表现为聚集分布,其余各阶段在各尺度下均趋于随机分布。(4)种群不同发育阶段不同尺度空间关联性不同,纯林样地中,幼苗与幼树在1—2 m小尺度上呈负相关,幼树与中龄树在8 m尺度呈正相关,其余各生长阶段之间在任意尺度上均无关联性;混交林样地中,幼树与中龄树在2—12 m尺度上表现为正相关,其余各生长阶段之间在各尺度上均无关联。在未来的森林经营中,应对不同发育阶段、不同生境的种群进行不同抚育措施,促进种群良性发育。     

Ovarian development status and population characteristics of Sogatella furcifera (Horváth) and Nilaparvata lugens (Stål): implications for pest forecasting

Migratory behaviour in insects correlates with reproductive development in females, and migration often occurs during the pre‐reproductive stage of adults. The relationship between ovarian development and population status of the white‐backed planthopper Sogatella furcifera (Horváth) and the brown planthopper Nilaparvata lugens (Stål) was evaluated. Females of both species were captured in rice fields and light traps and then dissected in double‐season rice‐farming regions of southern China. The ovarian development of S. furcifera and N. lugens was divided into five levels, following previous studies. The population statuses of both species were examined based on the ovarian development of female adults caught in rice paddies. The ovarian development in N. lugens females caught in light traps mostly ranged from level I to level II, whereas that in S. furcifera females caught in light traps mostly ranged from level I to level III. During peak immigration, ovarian development in N. lugens females was mainly at level II, whereas that in S. furcifera females was mainly at level II and sporadically at level III. During peak emigration, both S. furcifera and N. lugens showed level I ovarian development. The temporal dynamics of ovarian development in light trap catches revealed that (i) significant emigration and partial immigration periods occur in S. furcifera, with ovarian development mainly at level I and sporadically from level II to level III and (ii) numerous immigrants of N. lugens were detected during sedentary and local breeding periods. The temporal dynamics of ovarian development provides more information than does the paddy population. Thus, this study proposes another method for pest forecasting, which is more precise and efficient than conventional forecasting methods such as light trap catching and monitoring population dynamics in rice fields.     

Interacciones Planta‐Nodriza en Lophophora diffusa (Cactaceae) en un Desierto Subtropical de México1

One of the two species from the genus Lophophora is the false peyote Lophophora diffusa, an endemic cactus species of the xerophytic shrubland at Querétaro, México, considered threatened from illegal extraction due to its hallucinogenic and medicinal properties. We analyzed the spatial distribution of L. diffusa and its association with the locally dominant shrub species with the system SADIE (Spatial Analysis by Distance IndicEs). We also studied the principal microclimatic factors (light, temperature, and humidity) beneath the canopy of the possible nurse plant species. The use of SADIE is a new approach to study the mechanisms of spatial distribution. Lophophora diffusa and the shrub species presented an aggregated distribution with patches and gaps, as indicated by the values of I_a= 4.179 for L. diffusa and I_a= 1.660 for the vegetation. Lophophora diffusa was positively associated with the arboreal vegetation, particularly with Larrea tridentata and Acacia sororia, but was negatively associated with Celtis pallida and Myrtillocactus geometrizans. Microclimate evaluation indicated that C. pallida canopy significantly reduced radiation and temperature compared to the other species (L. tridentata, A.sororia y P. laevigata). We had expected L. diffusa to exhibit a positive spatial association with C. pallida; however, the reduction in light availability apparently limited seedling establishment of L. diffusa.     

Spatial genetic structure within two populations of a self-incompatible perennial,Chionographis japonica var.japonica (Liliaceae)

Intrapopulational spatial genetic structure was examined in two populations ofChionographis japonica var.japonica, a self-incompatible perennial, by spatial autocorrelation analysis of enzyme polymorphism. Although most spatial autocorrelation indices (Moran'sI) in the shortes distance class were significantly positive, most in the other distance classes did not significantly deviate from the values expected from random distributions of genotypes in both populations. This contrasts with a spatial genetic pattern previously reported for a population of the predominantly selfing congener,C. japonica var.kurohimensis, indicating that pollen-mediated gene flow highly impedes genetic substructuring within populations of outcrossingC. japonica var.japonica. Genetic similarity in very proximate distance found in outcrossingC. japonica var.japonica is probably due to restricted dispersal of seeds.     

Spatial correlograms of soil cover as an indicator of landscape heterogeneity

Soil cover, which is one of the most informative and integrative landscape factors, can be used for the analysis of landscape patterns. We studied the spatial autocorrelation (Moran's I) of raster format soil maps (1:10,000; 10 m pixel size) in 35 study areas representing all landscape regions in Estonia. The carbonate concentration of soils, volumetric soil moisture (%) and the depth of the groundwater table were taken into consideration in compiling a scale of contrast of 17 soil groups. We introduce a simple characteristic based on spatial correlograms: a half-value distance lag, h_I = 0.5—a distance where Moran's I drops below 0.5. Spatial autocorrelation decreased very rapidly in the case of heights with a very heterogeneous landscape composition, showing low values of h_I = 0.5 (<100 m in all 6 study areas). In uplands and depressions, the spatial autocorrelation also decreased relatively rapidly (h_I = 0.5 < 200 m). In most of the plains, coastal lowlands, sea islands and inland paludified lowlands, the values of Moran's I did decrease slowly with increasing lag, being >200 m in all forest and bog areas with complex topographical conditions due to the variety of glacial landforms and peatlands. All of the eight FRAGSTATS landscape metrics studied demonstrated significant correlations with h_I = 0.5, whereas five of them – Contrast Weighted Edge Density (CWED); Percentage of Like Adjacencies (PLADJ), Edge Density (ED), Patch Density (PD) and Mean Patch Area Distribution (AREA_MN) – had Spearman Rank Order Correlation values higher than 0.8. Landscapes with high ED, PD, and CWED values have a low autocorrelation: PD, ED, and CWED correlated negatively with h_I = 0.5. PD, ED, and CWED decreased and PLADJ increased with the power-law relationship with increasing h_I = 0.5. Spatial autocorrelation is lower in landscapes with complex structure and high contrast. The positive relationship with PLADJ indicates the same. Thus, spatial correlograms of potential landscape structure based on soil cover analysis can be used for the characterization of human-influenced landscape (land use) structure.     

Comparative study of movement patterns of Mastomys natalensis in irrigated rice and fallow fields in eastern Tanzania

A 2‐year capture–mark–recapture study was conducted to estimate home ranges and weekly travel distance of Mastomys natalensis (Smith 1834) in an irrigated rice ecosystem and fallow fields. We found that adults have larger home ranges than subadults in fallow fields but not in rice fields, indicating that fallow fields are more suitable for breeding. Travel distances were larger in rice fields, especially in the transplanting stage, during which rice fields are flooded and provide less food, causing movements into neighbouring fallow fields that then temporarily experience higher population density. A decrease in travel distance was observed in rice fields during the maturity stage, which can be explained by higher food availability and a more suitable, nonflooded situation. Movement of M. natalensis in rice‐fallow mosaic landscapes thus seems to be driven by food availability and flooding status of the rice fields, which can be attributed to land use practices.     

Expression of photosynthesis-related genes and their regulation by light during somatic embryogenesis in<Emphasis Type="Italic"> Daucus carota</Emphasis>

To clarify the spatial and temporal pattern of gene expression for photosynthesis-associated proteins during somatic embryogenesis in Daucus carota L., the localization of mRNAs for three genes, rbcL, Lhcb and por, was examined in dark-grown and light-irradiated somatic embryos by in situ hybridization. The three mRNAs were expressed in common in the mesophyll precursor cells of light-irradiated embryos at the late torpedo and plantlet stages, but characteristic expression patterns of each photosynthesis-related gene were also observed. Expression of rbcL mRNA first occurred throughout the embryo but gradually became localized in the mesophyll precursor cells and cortex during early embryogenesis. Localization of Lhcb mRNA in the mesophyll precursor cells and shoot apical meristem became clear in the early torpedo stage. Expression of Lhcb mRNA was not affected by light during early embryogenesis, but could be induced by light in the torpedo stage, suggesting that light-inducible expression of Lhcb mRNA arises within the torpedo stage. At the late torpedo stage, clear localization of por mRNA started in mesophyll precursor cells of the cotyledon in light-irradiated embryos. Greening potency of the embryo also appeared first at this stage. Therefore, greening and initial differentiation of photosynthetic tissues during somatic embryogenesis seem to be associated with coordinated expression of mRNA for rbcL, Lhcb and por in late torpedo-shaped embryos.Abbreviations DIG Digoxigenin - Lhcb3 Gene encoding a type-III light-harvesting chlorophyll a/b-binding protein of photosystem II - LHCII Light-harvesting chlorophyll a/b-binding protein of photosystem II - POR Protochlorophyllide oxidoreductase - rbcL Gene encoding the large subunit of Rubisco - Rubisco Ribulose-1,5-bisphosphate carboxylase/oxygenase     

Photosynthetic and biochemical activities in flag leaves of a newly developed superhigh-yield hybrid rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>) and its parents during the reproductive stage

Responses of net photosynthetic rates to intercellular CO₂ concentration (P _n/C _i curves) and photochemical characteristics were investigated in flag leaves of newly developed superhigh-yield hybrid rice (Oryza sativa L.) LiangYouPeiJiu (LYPJ) and its maternal PeiAi64S (PA64S) and paternal WuMang9311 (WM9311) lines grown in the field during the reproductive stage. The results showed that photosynthetic functions, such as the electron transport activities of photosystems and photophosphorylation, assessed in vivo from P _n/C _i curves under field conditions declined more or earlier than those obtained in vitro. The degradation of polypeptides of thylakoid membranes was slower than those for P _Ca=360 (light-saturated net photosynthetic rate measured at 360 μmol mol⁻¹) and CE (carboxylation efficiency, obtained from the initial slope of the P _n/C _i curve). The initial inhibition of the PSII electron transport and oxygen-evolving activity induced by senescence occurred before the degradation of the oxygen-evolving complex. In comparison, LYPJ had intermediate photosynthetic functions in the early stage of leaf development, but greater photochemical activities in the mid and late stages. WM9311 showed a similar pattern of changes but lower values, and PA64S had higher values in the early stage but showed a faster rate of senescence than LYPJ. These findings implied that the hybrid LYPJ demonstrated intermediate photosynthetic activities between its parents in the early stage of leaf development, whereas it had higher photosynthetic activities than its parents in the mid and late stages, which may be responsible for its high yield.     

Effect of the Juvenile Hormone Analog,Fenoxycarb on Termination of Reproductive Diapause in Scotinophara lurida (Burmeister) (Heteroptera: Pentatomidae)

This study was carried out to investigate a role of the juvenile hormone analog, fenoxycarb (FC) in terminating the reproductive diapause in the black rice bug, Scotinophara lurida (Burmeister) (Heteroptera: Pentatomidae). FC-treated adults collected at different dates developed their ovaries sooner than untreated ones. Topical application of FC accelerated diapause termination of field-collected S. lurida in which FC-treated adults exhibited early oviposition. In FC-treated males, development of their accessory glands and testes accelerated appeared to be prompted. These results suggest that various physiological overwintering traits related to diapause in this species may in part be under gonadotrophic endocrine control.     

A Study of Genetic Structure of Stephania yunnanensis (Menispermaceae) by DALP

Genetic diversity and genetic structure within and among ten populations of Stephania yunnanensis H. S. Lo and three populations of S. epigaea H. S. Lo from Yunnan province were evaluated by direct amplification of length polymorphism (DALP) markers. Five primer groups were screened, and a total of 287 DNA fragments were amplified, among which 266 were polymorphic, averaging 53.2 polymorphic bands per primer group in S. yunnanensis. The percentage of polymorphic bands of S. yunnanensis was 92.68% at the species level and 61.92% within the ten populations sampled. At the species level, the observed number of alleles (N _a) was 1.9268 and the effective number of alleles (N _e) was 1.5933; Nei’s gene diversity (H) was 0.3414; Shannon’s information index (I) was 0.5057. At the population level, N _a = 1.6192, N _e = 1.4001, H = 0.2298, and I = 0.3401. Total gene diversity of S. yunnanensis was 0.3419. Gene diversity within population was 0.2298, coefficient of gene differentiation was 0.3278, and estimated gene flow was 1.0254. The results indicated that the genetic differentiation was relatively higher among populations of S. yunnanensis. DALP markers were an informative and useful method for assaying genetic diversity and authenticating species of Stephania. These data could provide basic molecular evidence for establishing a reasonable strategy for protecting and exploiting the resource of S. yunnanensis.     

Validation of Candidate Reference Genes for the Accurate Normalization of Real-Time Quantitative RT-PCR Data in Rice During Seed Development

Rice seed, a natural storage organ for starch and protein, is also an ideal bioreactor for the production of valuable proteins. Increasingly, studies focused on rice have tried to determine the functions of its genes and also to improve its yield and quality. Real-time RT-PCR is the best available choice at present for gene expression analysis due to its accuracy, sensitivity, and reproducibility. The right choice of reference genes for normalization, however, is a critical precondition for reliable results. In this study, the expression stabilities of nine commonly used housekeeping genes in rice were carefully assessed using the software geNorm. Our results showed that eIF-4a and ACT1 were the most suitable reference genes among almost all the tested samples from two rice varieties, including different temporal and spatial-specific tissues, especially in seeds at different developmental stages. In contrast, 18S and 25S rRNAs, two common reference genes, were found to have the least stable expression. Moreover, it is necessary to use multiple suitable reference genes together for normalization to get a more reliable result in temporal and spatial expression analysis during rice seed development. The validated reference genes were further relied when used to quantify the expression of several genes of interest during rice seed development.