Evaluation of the Characteristics of Virus-free Sweetpotato and Its Use in Hybridization

Application of virus-free sweetpotato is a breakthrough of sweetpotato production. Several popular varieties were adopted to study their yield increase potential and their use in breeding in this research. The results showed that after virus-elimination, all varieties had a yield increase ranging from 14.95% - 91. 61% and marketable quality improvement, despite the changes in location and season. The changes of dry matter content and disease resistance were not significant. Virus-free sweetpotato performed vigorous vine growth and strong dry matter assimilation ability. Virus-free parents had a slightly high seed setting percentage, but its flowering ability and performance of off-springs were similar to that of its check plants.

Detection of Ratoon Stunting Disease in Virus-free Seedcane via Real-time Fluorescence Quantitative PCR

This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease （RSD） in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from six plants of sugarcane ROC22, which had been confirmed RSD-positive by detecting the sugarcane juice, by employing the sugarcane seedlings production protocol. Real-time fluorescence quantitative PCR was used to detect RSD pathogens in tissue culture sam- pies. The results showed that target fragment of RSD pathogens was not found in all 10 samples in real-time fluorescence quantitative PCR, with the Ct values of 37 - 39. The healthy tissue culture sugarcane seedlings do not carry RSD pathogens, indicating that adopting healthy seedcane seedlings production technique could thoroughly get rid of RSD pathogens.

A Preliminary Study on Tissue Culture and Rapid Propagation of Virus-free Plantlets of Benihoppe Strawberry

[ Objective] This study aimed to investigate the tissue culture and rapid propagation techniques of Benihoppe strawberry. [ Method] Shoot tips of new stolons of Benihoppe strawberry were used as experimental materials to analyze the effects of media type, cytokinin type and concentration, carbon source type and concentration, and light intensity on tissue culture and propagation of Benihoppe strawberry. [ Result] MS was the optimal media for plantlet propagation. In the media containing 1.2 ing/L BA （with addition of 0.1 mg/L NAA）, fresh weight, dry weight and propagation coefficient of strawberry plantlets reached the maxi- mum, which were 2.259 g, 0. 221 g and 12.4, respectively. The optimal carbon source was 30 g/L sucrose, and the optimal light intensity was 1 600 lx. [ Conclu- sion] The best media for tissue culture and rapid propagation of Benihoppe strawberry was MS ＋ BA 1.2 mg/L ＋ NAA 0. 1 mg/L ＋ sucrose 30 g/L ＋ agar 8 g/L. This study provided scientific basis for large-scale production of Benihoppe strawberry.

Expression of Invertase Genes in Virus-free Sugarcane

Virus-free sugarcane seedlings have improved biomass and sucrose content compared with ordinary seedlings, and sucrose invertases are key enzymes regulating sugarcane growth and sucrose accumulation. In this study, the differences in the expression levels of 3 invertase genes, CWI, SAI and NI, between virus- free and ordinary sugarcane seedlings were analyzed. Compared with ordinary sugarcane plants, the expression of CWI was mainly up-regulated in immature leaves and stems at elongation stage and leaves and immature internodes at maturation stage, and especially, greatly up-regulated in immature interuedes at maturation stage of virus-free plants. The expression of SAI and NI were mainly up-regnlated in leaves and immature internedes of virus-free plants at maturation stage, which might be beneficial to sugar accumulation and rapid utilization of monosaccharide in the stalks of virus-free plants. It is further indicated that virus-free treatment could significantly improve the expression of sucrose invertases at late growth period, and might facilitate the increase of plant biomass.

Study on Virus-free Breeding Technology of Potato Microtuber

It is the simplest,fastest and effective way to improve the yield level of potato by selecting high quality virus-free seed potato for field production and maintaining the original characters of virus-free minituber. Microtuber is featured by small volume,no pathogen,convenient storage and transportation,which can be induced in summer with high temperature and humidity,and overcome a series of problems such as poor survival rate of transplanted test-tube plantlets,and easy pollution of cutting tips. Therefore,it can replace conventional test-tube plantlets directly for reproduction. Meantime,if minitubers are sown ahead of time using cold frame,the growth stage of potato minituber will avoid the occurrence period of aphids. The use of gauze cover for insect prevention has good effect on maintaining the excellent characters of microtuber. The yield and quality of virus-free potato are greatly improved by means of virus-free culture of shoot tip seedlings,subculture propagation of virus-free seedlings,induction of microtubers by dark culture,cultivation of breeder’s seeds in greenhouse and screenhouse to preserve the characters of potato.

Correlations between delayed fluorescence of chlorophyll, metabolism and yield of plants. III. Influence of viral infection on field plants and new technology of clone selection of virus-free planting potato

At the end of potato plants vegetation virus in-fection induced both decrease in maximum amplitude of delayed fluorescence maximal amplitude and increase half time of its decrease, as well as reduction in the amount of stems, plants’ height and assimilation area surface, yield, acceleration of plants development and their early die-off. The differences of DF pa-rameters and yields between strongly and weakly infected plants increase in case of a combined virus infection. In industrial test of the selection of virus-free planting potato by the use of DF parameter, a rise in the yield and de-crease degree of viral infection of crops was obtained.

Barley FASCIATED EAR genes determine inflorescence meristem size and yield traits

In flowering plants,the inflorescence meristem(IM)provides founder cells to form successive floral meristems,which are precursors of fruits and seeds.The activity and developmental progression of IM are thus critical for yield production in seed crops.In some cereals,such as rice(Oryza sativa)and maize(Zea mays),the size of undifferentiated IM,which is located at the inflorescence apex,is positively associated with yield traits such as spikelet number.However,the relationship between IM size and yieldrelated spike traits remains unknown in the Triticeae tribe.Here we report that IM size has a negative correlation with yield traits in barley(Hordeum vulgare).Three FASCIATED EAR(FEA)orthologs,HvFEA2,HvFEA3,and HvFEA4,regulate IM size and spike morphogenesis and ultimately affect yield traits.Three HvFEAs genes are highly expressed in developing spikes,and all three loss-of-function mutants exhibit enlarged IM size,shortened spikes,and reduced spikelet number,which may lead to reduced grain yield.Natural variations identified in HvFEAs indicate selection events during barley domestication.We further reveal that HvFEA4,as a transcription factor,potentially targets multiple pathways during reproductive development,including transcriptional control,phytohormone signaling,and redox status.The roles of barley FEA genes in limiting IM size and promoting spikelet formation suggest the potential of increasing yield by manipulating IM activity.

B Class Floral Homeotic Genes are Involved in the Petal Identity and Flower Meristem Determinations in Chrysanthemum morifolium

Chrysanthemum morifolium,an ornamental crop with diverse forms of inflorescence,is a good model for studying flower development in Asteraceae.However,the genetic background is complex and the mechanisms of regulating flower development are still unclear.Here,we identified two natural mutant lines of chrysanthemum and named them M1 and M2 according to the severity of the phenotype.Both lines showed defects in petal identity,and the petals of the M1 line had a mild phenotype:partially loss of petal identity and conversion of petals into green,leaf-like organs.The M2 line had severe phenotypes:in addition to severe petal defects,secondary inflorescences were produced in the capitulum to replace the normal ray and disc florets,which indicated a transformation of a flower meristem into an inflorescence meristem.Transcriptome sequencing of WT and M2 inflorescences was performed and found altered expression of floral organ development A,B and E class genes,where B and E class genes were significantly down-regulated.qRT-PCR analysis in both M1 and M2 lines revealed that the expression of three chrysanthemum class B genes CmAP3.1,CmAP3.2 and CmPI,was negatively correlated with phenotypic severity.This suggests that class B genes in chrysanthemum not only have conserved functions in determining petal identity but also were involved in the determinacy of the flower meristem.This study provides insights into the functions of class B genes in flower development,and is informative for dissecting the molecular mechanisms of flower development in chrysanthemum.

Steps To ward HIV Cure Two more HIV-infected patients appear to be virus-free after bone-marrow transplants

THIS year＇s International AIDS Society Conference on HIV Pathogenesis, Treatment and Preven-／tion, held in Kuala Lumpur, Malaysia, made major headlines when Timothy Hendch, an American doctor, announced that two more cancer patients may have been cured of HIV after receiving bone-marrow transplants to treat lymphoma. Both patients had been taking retroviral medication, and continued to do so after the transplants as their viral levels sank until doctors were unable to find any traces of HIV in the patients＇ blood.

激素调控植物分枝/分蘖的研究进展

分枝/分蘖是植物株型的一个重要特征,也是腋芽起始和芽生长的结果,对经济作物的种子产量以及牧草产量均具有决定性作用。多种激素及其互作效应在植物分枝/分蘖的发生和生长发育过程中起着关键的调控作用,且环境因素也是通过改变植物体内激素含量及其平衡调控分枝的。本研究综述了多种激素对植物分枝/分蘖调控机制的多个方面,包括生长素、细胞分裂素、独脚金内酯、油菜素内酯、脱落酸和赤霉素、乙烯、茉莉酸及不同激素信号相互作用形成的复杂调控网络,旨在为利用激素调控机制培育具有理想株型的高产新作物品种奠定基础。同时分析了激素机制调控植物分枝/分蘖的现存问题,并展望了激素调控植物分枝/分蘖的研究方向,以期为通过激素调控改良作物株型提供理论依据。

The AGL6-like gene OsMADS6 regulates floral organ and meristem identities in rice

Although AGAMOUS-LIKE6 (AGL6) MADS-box genes are ancient with wide distributions in gymnosperms and angiosperms, their functions remain poorly understood. Here, we show the biological role of the AGL6-1ike gene, OsMADS6, in specifying floral organ and meristem identities in rice (Oryza sativa L.). OsMADS6 was strongly ex- pressed in the floral meristem at early stages. Subsequently, OsMADS6 transcripts were mainly detectable in paleas, lodicules, carpels and the integument of ovule, as well as in the receptacle. Compared to wild type plants, osmads6 mutants displayed altered palea identity, extra glume-like or mosaic organs, abnormal carpel development and loss of floral meristem determinacy. Strikingly, mutation of a SEPALLATA (SEP)-like gene, OsMADS1 (LHS1), enhanced the defect of osmads6 flowers, and no inner floral organs or glume-like structures were observed in whorls 2 and 3 of osmadsl-z osmads6-1 flowers. Furthermore, the osmadsl-z osmads6-1 double mutants developed severely indetermi- nate floral meristems. Our finding, therefore, suggests that the ancient OsMADS6 gene is able to specify "floral state" by determining floral organ and meristem identities in monocot crop rice together with OsMADS1.

Determination of the Photoperiod-Sensitive Inductive Phase in Maize with Leaf Numbers and Morphologies of Stem Apical Meristem

It is vital to determine the effective photoperiods of maize for making full use of tropical germplasm, which is the foundation for determining the effect of latitude and planting date on the development of photoperiod-sensitive maize cultivars. The objective of this study is to determine the photoperiod-sensitive inductive phase using reciprocal transfer between long- day （LD） （15 h d^-1） and short-day conditions （SD） （9 h d^-1）. For Huangzao 4 and CML288, days to tassel and pollen shedding were recorded, and stem apical meristems （SAM） were observed by a laser scanning confocal microscope. The results show that the seedlings are insensitive to photoperiod when they are very young （juvenile）. However, after this period, LD delays flowering and increases the leaf numbers below the inflorescence, and the length of the interval of the photoperiod-sensitive inductive phase is longer under LD conditions than under SD conditions. Transferred from SD to LD, plants show a sudden decrease in leaf numbers once sufficient SD has been received for flower commitment. While transferred from LD to SD, plants have a continuous increase in leaf numbers during the photoperiod sensitive inductive phase under LD conditions. At the same time, when plants are competent to flowers, the obvious morphology is the elongation of maize SAM. There is an obvious variance of the photoperiod sensitive phase under LD and SD conditions in different maize.

AtCDC5 regulates the G2 to M transition of the cell cycle and is critical for the function of Arabidopsis shoot apical meristem

作为一个房间周期管理者， Myb 相关的 CDC5 蛋白质被报导为在酵母和动物的房间周期的 G2 阶段必要，但是很少在植物对它的功能被知道。这里，我们在 Arabidopsisthaliana 报导 CDC5 基因的功能的描述。Arabidopsis CDC5 (AtCDC5 ) 主要与高细胞分裂活动在纸巾被表示，并且在整个胚胎形成的全部过程被表示。AtCDC5 loss-of-functionmutant 是胚胎的致命。以便调查 AtCDC5 体内的函数，我们 AtCDC5 的表达式被 RNA 干扰在减少的 generatedAtCDC5-RNAi 植物。我们发现到 M (G2/M ) 阶段转变的 G2 在 AtCDC5-RNAi 植物被影响，并且 thatendoreduplication 被增加。另外，射击顶端分生组织(SAM ) 的维护功能在 AtCDC5-RNAi 植物被扰乱，在哪个 WUSCHEL (WUS )-CLAVATA (CLV ) 和射击无分裂组织(STM ) 小径被损害。原位杂交分析证明 STM 的表示极大地在 AtCDC5-RNAi 植物的射击顶端细胞被减少。而且， cyclinB1 或 Histone4 被发现当时，在一些这些房间被表示 STMwas 的抄本无法发现。这些结果建议 AtCDC5 为 G2/M 阶段转变是必要的并且可以由控制 STM 和 WUS 的表示调整 SAM 的功能。

The GhREV transcription factor regulate the development of shoot apical meristem in cotton(Gossypium hirsutum)

Background:Manual topping is a routine agronomic practice for balancing the vegetative and reproductive growth of cotton(Gossypium hirsutum)in China,but its cost-effectiveness has decreased over time.Therefore,there is an urgent need to replace manual topping with new approaches,such as biological topping.In this study,we examined the function of Gh REV transcription factors(a classⅢhomeodomain-leucine zipper family,HD-ZIPⅢ)in regulating the development of shoot apical meristem(SAM)in cotton with the purpose of providing candidate genes for biological topping of cotton in the future.Results:We cloned four orthologous genes of At REV in cotton,namely Gh REV1,Gh REV2,Gh REV3,and Gh REV4.All the Gh REVs expressed in roots,stem,leaves,and SAM.Compared with Gh REV1 and Gh REV3,the expression level of Gh REV2 and Gh REV4 was higher in the SAM.However,only Gh REV2 had transcriptional activity.Gh REV2 is localized in the nucleus;and silencing it via virus-induced gene silencing(VIGS)produced an abnormal SAM.Two key genes,Gh WUSA10 and Gh STM,which involved in regulating the development of plant SAM,showed about 50%reduction in their transcripts in VIGS-Gh REV2 plants.Conclusion:Gh REV2 positively regulates the development of cotton SAM by regulating Gh WUSA10 and Gh STM potentially.

The Wheat Plastochron Mutant, fushi-darake, Shows Transformation of Reproductive Spikelet Meristem into Vegetative Shoot Meristem

In wheat plants at the vegetative growth stage, the shoot apical meristem (SAM) produces leaf primordia. When reproductive growth is initiated, the SAM forms an inflorescence meristem (IM) that differentiates a series of spikelet meristem (SM) as the branch. The SM then produces a series of floret meristem (FM) as the branch. To identify the mechanisms that regulate formation of the reproductive meristems in wheat, we have investigated a leaf initiation mutant, fushi-darake (fdk) which was developed by ion beam mutagenesis. The morphological traits were compared in wild type (WT) and fdk mutant plants grown in the experimental field. WT plants initiated leaves from SAM at regular intervals in spiral phyllotaxy, while fdk plants had 1/2 alternate phyllotaxy with rapid leaf emergence. The fdk plants have increased numbers of nodes and leaves compared with WT plants. The time interval between successive leaf initiation events (plastochron) was measured in plants grown in a growth chamber. The fdk plants clearly show the rapid leaf emergence, indicating a shortened plastochron. Each tiller in fdk plants branches at the upper part of the culm. The fine structure of organ formation in meristems of fdk plants was examined by scanning electron microscopy (SEM). The SEM analysis indicated that fdk plants show transformation of spikelet meristems into vegetative shoot meristems. In conclusion, the fdk mutant has a heterochronic nature, i.e., both reproductive and vegetative programs were simultaneously in operation during the reproductive phase, resulting in a shortened plastochron and transformation of reproductive spikelets into vegetative shoots.

Floral Organogenesis and Ring Meristem in <i>Phytolacca</i>

To further study the floral organogenesis and discussing the floral origin of Phytolacca, the procedures of floral organogenesis were observed in Phytolacca esculenta and Phytolacca zhejiangensis. The results showed that the floral organogenesis was consistent in Phytolacca. Their sepals were 2/5 helix, and with counter-clockwise and clockwise, usually the first sepal located at non-median of abaxial side. The first sepal of Phytolacca esculenta was initiated at non-median of adaxial side. There was no evident relationship between sepal and stamen initiating position, and the stamens initiated on ring meristem, they initiated approximately at the same time, and when the androecium member was numerous, they initiated centrifugally, the outer stamen initiated irregularly. Carpel initiated alternately with inner stamens. And the carpels connected by septum, if the septum grew more, the carpel was syncarpous at morphology, otherwise the carpel was apocarpous at morphology. So the syncarpous and the apocarpous have no successively relationship on evolution. Ovule initiated inside the carpel and opposite to carpel. Androecium, carpel and ovule initiated at ring meristem.

Generation of Virus Free Potato Plantlets through Meristem Culture and Their Field Evaluation

Different aspects of micropropagation through meristem culture for the production of virus indexed source plants, in vitro tuberization and field evaluation of the in vitro regenerated plants were studied on four commercial cultivars of potato (Solanum tuberosum L.) viz., Diamant, Cardinal, Shilbilati and Lalpakri. The investigation was conducted at Rajshahi, Bangladesh from December 2010 to March 2012 to produce virus-free potato plantlets through meristem culture, shoot multiplications with root induction as well as their acclimatization and evaluation of morphological characters and tuber yield under field condition. Shoot tips of 25 - 30 day old field-grown plants of above mentioned four cultivars were used for meristem isolation. After isolation, meristems of these varieties of potato were cultured on “M” shaped filter paper bridge in Murashige and Skoog (MS) liquid medium. Four different treatments of media formulations viz. 0.1 mg/L KIN + 0.1 mg/L GA3, 0.1 mg/L KIN + 0.5 mg/L GA3, 0.5 mg/L KIN + 0.1 mg/L GA3 and 0.5 mg/L KIN + 0.5 mg/L GA3 were used as plant growth regulators. From these formulations MS + 0.1 mg/L KIN + 0.5 mg/L GA3 was found to be the best for the primary establishment of meristem culture. The primarily established meristems were subcultured on to MS semisolid basal medium supplemented with four different treatment combinations of hormones viz. 0.5 mg/L BA + 1.0 mg/L IBA;0.1 mg/L KIN + 0.1 mg/L GA3;0.5 mg/L BA + 0.5 mg/L GA3 and 0.5 mg/L KIN + 0.5 mg/L GA3 were used to identify the suitable media compositions for shoot proliferation. Results showed that out of these four media treatments the formulation 0.5 mg/L BA + 0.5 mg/L GA3 was found to be the best suitable for shoot generation. Among the four cultivars of potato higher frequency of shoot proliferation (number of shoots/explant and longest shoot length) was observed in Diamant, though the highest shoot formation (76%) was recorded in Cardinal. Virus free in vitro grown potato plantlets were obtained through DAS-ELISA test and used substantially for micro-propagation. After gradual acclimatization of rooted plantlets of four potato cultivars, they were transferred into the field for cultivation and established successfully. It was observed from the field study of in vitro meristem-derived plantlets that there were no virus-affected plants. The virus-free exotic varieties were much superior in all vegetative attributes and yield compared to those of indigenous varieties with producing potato plants of normal height. In contrast, the indigenous varieties took a longer time to tuber initiation and maturity, lower plant height and number of leaves per plant, a higher number of tubers but a lower amount of tuber weight per plant, and poorer tuber grade than the exotic varieties. However, the variety Cardinal exposed the best performances in the context of survival percentage of plantlets (90%), days to tuber initiation (DTI), a number of leaves per plant (NL), tuber weight per plant (343.40%) and the percentage of rich tuber grade.

Efficient Regeneration System for Genetic Transformation of Mulberry (<i>Morus indica</i>L. Cultivar S-36) Using <i>in Vitro</i>Derived Shoot Meristems

Shoot meristems used for the study were exercised from the in vitro regenerated shoots cultured on MS medium supplemented with 0.5 mg/L of BAP for multiplication. The sensitivity of the in vitro regenerated was studied using shoot meristems of 0.5 cm. Shoot meristems were cultured on medium containing 10-100 mg/l kanamycin to determine the concentration that was lethal for multiple shoot induction and root induction. The response of shoot multiplication decreased (66.2%-6.2%) as the concentration of kanamycin increased (10.0-70.0 mg/L) with complete inhibition of shoot proliferation at 100 mg/L kanamycin. The rooting phase was very sensitive to kanamycin compared to shoot multiplication. The percentage of shoots that rooted decreased (53.8%-4.8%) with increase in the concentration of kanamycin (10.0-70.0 mg/l) on IBA and 2,4-D supplemented medium. For transformation studies, the shoot tips that were infected with Agrobacterium strain were placed on selection medium containing MS medium with 0.5 mg/L BAP and 100 mg/L kanamycin and scored for the putative transformed shoots. An average of 62.2% of shoot tips developed shoot buds from the base and the shoots reached a length of 0.5-1.0 cm at the end of 30 days of culture on the selective medium in comparison to control which showed no response. An average of 66.7% of the regenerated plants showed GUS expression on selection medium where 43.2% and 65% of GUS expression was recorded in the leaves and callus. Leaves and callus induced from the controls did not show GUS activity. Stable integration of nptII gene with the genomic DNA from these transformed plants was confirmed through PCR analysis. Our result presents an efficient regeneration system using in vitro derived shoot meristems for Agrobacterium mediated gene transfer.

The Effects of Lead on the Meristem of Wheat Seedlings

The ultrastructure of apical meristem cells was studied in Triticum aestivum L. cv. “Trizo” seedlings grown on soil without or enriched with selenium and survived 14 days’ stress caused by lead pollution in the soil. The soil treatments: control—the original soil;(Pb1)—50 mg&middot;kg&minus;1;(Pb2)—100 mg&middot;kg&minus;1;(Pb1 + Se1) —0.4 mg&middot;kg&minus;1 Se added to Pb1 treated soil;(Pb1 + Se2)—0.8 mg&middot;kg&minus;1 Se added to Pb1 treated soil;(Pb2 + Se1)—0.4 mg&middot;kg&minus;1 Se added to Pb2 treated soil;(Pb2 + Se2)—0.8 mg&middot;kg&minus;1 Se added to Pb2 treated soil were used. Light and other conditions were optimal for plant growth. A distinctive feature of the cells of the apical meristem of control plants was the absence of nuclear membranes. Proplastids were membrane vesicles 1 - 2 microns in diameter, filled with contents of varying degrees of density, from membrane vesicles containing only plastid DNA up to a fully formed structure of proplastids. In (Pb1)-plants, the amount of cytoplasmic ribosomes and proplastids in the meristematic cells was less than in the control. The structure of the forming proplastids was almost the same as that of the control plants. Signs of degradation of meristematic proplastids, such as a decrease of their diameter, observed in (Pb2)-plants. The introduction of selenium into lead contaminated soil increased the accumulation of Pb in plants, especially in the roots and apical meristem. In (Pb1 + Se1)-, (Pb1 + Se2)-, (Pb2 + Se1)-, and (Pb2 + Se2)-plants, the number of cytoplasmic ribosomes in meristematic cells increased, which indirectly indicates an increase in protein synthesis. Based on our concept about the formation (assembly) of proplastids in the cells of the apical meristem, we believe that toxic agents, such as lead, which inhibit the development of proplastids into chloroplasts in mesophyll cells, act on apical meristem cells at the stage when plastid DNA is replicated in the cytoplasm and is not yet surrounded by a plastid membrane.

Genetic and environmental control of rice tillering

Increasing tiller number is a target of high-yield rice breeding. Identification of tiller-defect mutants and their corresponding genes is helpful for clarifying the molecular mechanism of rice tillering. Summarizing research progress on the two processes of rice tiller formation, namely the formation and growth of axillary meristem, this paper reviews the effects of genetic factors, endogenous hormones, and exogenous environment on rice tillering, finding that multiple molecular mechanisms and signal pathways regulating rice tillering cooperate rice tillering, and discusses future research objectives and application of its regulatory mechanism. Elucidation of theis mechanism will be helpful for breeding high-yielding rice cultivars with ideal plant type via molecular design breeding.