Plant Chemical Defenses against Insect Herbivores—Using theWild Tobacco as a Model

The Nicotiana genus, commonly known as tobacco, holds significant importance as a crucial economic crop. Confrontedwith an abundance of herbivorous insects that pose a substantial threat to yield, tobacco has developed adiverse and sophisticated array of mechanisms, establishing itself as a model of plant ecological defense. Thisreview provides a concise overview of the current understanding of tobacco’s defense strategies against herbivores.Direct defenses, exemplified by its well-known tactic of secreting the alkaloid nicotine, serve as a potent toxinagainst a broad spectrum of herbivorous pests. Moreover, in response to herbivore attacks, tobacco enhancesthe discharge of volatile compounds, harnessing an indirect strategy that attracts the predators of the herbivores.The delicate balance between defense and growth leads to the initiation of most defense strategies only after aherbivore attack. Among plant hormones, notably jasmonic acid (JA), play central roles in coordinating thesedefense processes. JA signaling interacts with other plant hormone signaling pathways to facilitate the extensivetranscriptional and metabolic adjustments in plants following herbivore assault. By shedding light on these ecologicaldefense strategies, this review emphasizes not only tobacco’s remarkable adaptability in its natural habitatbut also offers insights beneficial for enhancing the resilience of current crops.

Development of a stable attenuated double-mutant of tobacco mosaic virus for cross-protection

Tobacco(Nicotiana tabacum)and tomato(Solanum lycopersicum)are two major economic crops in China.Tobacco mosaic virus(TMV;genus Tobamovirus)is the most prevalent virus infecting both crops.Currently,some widely cultivated tobacco and tomato cultivars are susceptible to TMV and there is no effective strategy to control this virus.Cross-protection can be a safe and environmentally friendly strategy to prevent viral diseases.However,stable attenuated TMV mutants are scarce.In this study,we found that the substitutions in the replicase p126,arginine at position 196(R^(196))with aspartic acid(D),glutamic acid at position 614(E^(614))with glycine(G),serine at position 643(S^(643))with phenylalanine(F),or D at position 730(D^(730))with S,significantly reduced the virulence and replication of TMV.However,only the mutation of S^(643) to F reduced the RNA silencing suppression activity of TMV p126.A double-mutant TMV-E614G-S643F induced no visible symptom and was genetically stable through six successive passages in tobacco plants.Furthermore,our results showed that TMV-E614G-S643F double-mutant could provide effective protection against the wild-type TMV infection in tobacco and tomato plants.This study reports a promising mild mutant for cross-protection to control TMV in tobacco and tomato plants.

Evaluation of Protector 700 Wd in Controlling Tobacco Pests in Tanzania

In Tanzania, tobacco is an important cash crop benefiting most farmers and stakeholders. However, the crop is characterized by a low yield per unit area. Several factors are responsible for the low yield and quality of tobacco, among which are damages caused by the insects and other pests. Major insect pests of tobacco are cutworms (Agrotis spp), budworms (Heliothis virescens (F.)), (Lepidoptera Noctuidae) and aphids (Myzus persicae, Aphis tabaci) (Homoptera: Aphididae). These pests adversely affect crop growth, yield and leaf quality. In order to reduce insect attack in tobacco, applying pesticides is a critical management practices. In Tanzania, various pesticides have been approved for use in controlling the pests. Currently, farmers are using Bamethrine 2.5 EC to control the pests. The Pesticide named Protector 700 WG with an active ingredient imidacloprid 700 g/kg is a new product introduced in Tanzania during the 2021/22 crop season. Research trials were conducted in three research sites, namely Ulowa-Kahama, Tumbi-Tabora and Mtanila-Chunya, on Protector 700 WG efficacy in different rates, 15 g, 20 g, and 25 g each dissolved in 20 litres of water. The Protector 700 Wg was compared with the standard pesticide (Bamethrin 2.5 EC). Absolute control was also inclusive where the plot had no application of pesticides. The results showed that Protector 700 WG at all rates outweigh the commonly used pesticide. The difference in the number of pests between Protector 700 WG and Bamethrin 2.5 EC was significantly at P < 0.005 also, in dry and green weights, the yield was significantly different at P < 0.005. However, when yield and number of insects were compared among the rates of protector, there was no significant difference. From these results, farmers are advised applying Protector 700 WG at the rate of 0.15 kg/ha by dissolving it in 200 litres of water.

Application of Sargassum fusiforme(Harv.)Setch.Extract in Cigarettes

[Objectives]This study was conducted to investigate the application of Sargassum fusiforme(Harv.)Setch.in cigarettes.[Methods]Tobacco-specific nitrosamines in the smoke of cigarettes added with ethanol extract of S.fusiforme were determined,and the compounds related to the aroma of S.fusiforme were identified by flavor-smelling experiment.[Results]With the addition of the ethanol extract of S.fusiforme,the decrease in the total amount of four tobacco-specific nitrosamines in mainstream cigarette smoke reached 16.42%.The results of the flavor-smelling experiment showed that the aroma of S.fusiforme might be related to(R)-5,6,7,7A-tetrahydro-4,4,7A-trimethyl-2(4H)-benzofuranone,glycerol,ethyl palmitate,methyl palmitate,ethyl linoleate,methyl(Z,Z,Z)-9,12,15-octadecatrienoate,ethyl(Z,Z,Z)-9,12,15-octadecatrienoate,phytol,and tetradecanoic acid.[Conclusions]The ethanol extract of S.fusiforme has the potential function of reducing the content of tobacco-specific nitrosamines in smoke and improving the taste of cigarettes.

Lateral root elongation in maize is related to auxin synthesis and transportation mediated by N metabolism under a mixed NO_(3)^(–) and NH_(4)^(+) supply

A mixed nitrate (NO_(3)^(–)) and ammonium (NH_(4)^(+)) supply can promote root growth in maize (Zea mays),however,the changes in root morphology and the related physiological mechanism under different N forms are still unclear.Here,maize seedlings were grown hydroponically with three N supplied in three different forms (NO_(3)^(–)only,75/25 NO_(3)^(–)/NH_(4)^(+)and NH_(4)^(+)only).Compared with sole NO_(3)^(–)or NH_(4)^(+),the mixed N supply increased the total root length of maize but did not affect the number of axial roots.The main reason was the increased total lateral root length,while the average lateral root (LR) length in each axle was only slightly increased.In addition,the average LR density of 2nd whorl crown root under mixed N was also increased.Compared with sole nitrate,mixed N could improve the N metabolism of roots (such as the N influx rate,nitrate reductase (NR) and glutamine synthase (GS)enzyme activities and total amino content of the roots).Experiments with exogenously added NR and GS inhibitors suggested that the increase in the average LR length under mixed N was related to the process of N assimilation,and whether the NR mediated NO synthesis participates in this process needs further exploration.Meanwhile,an investigation of the changes in root-shoot ratio and carbon (C) concentration showed that C transportation from shoots to roots may not be the key factor in mediating lateral root elongation,and the changes in the sugar concentration in roots further proved this conclusion.Furthermore,the synthesis and transportation of auxin in axial roots may play a key role in lateral root elongation,in which the expression of ZmPIN1B and ZmPIN9 may be involved in this pathway.This study preliminarily clarified the changes in root morphology and explored the possible physiological mechanism under a mixed N supply in maize,which may provide some theoretical basis for the cultivation of crop varieties with high N efficiency.

Advances in Research on Synthetic Microbial Communities

The synthetic microbial community is a synthetic microbial system co-cultured with multiple species, which has the characteristics of clear composition and strong controllability. Compared with a single colony, it can achieve more complex functions and adapt to the changing environment more easily, so as to meet a wide range of needs. In this paper, the contents and concepts of microbial community and synthetic microbial community are briefly introduced, the principles that should be followed in the construction of microbial community are expounded, the methods and mathematical models used in the construction of synthetic microbial community are introduced, and the applications of synthetic microbial community in various fields are summarized. Finally, the challenges in the research of synthetic microbial communities are briefly described.

A dicistrovirus increases pupal mortality in Spodoptera frugiperda by suppressing protease activity and inhibiting larval diet consumption

Understanding interactions between viruses and their hosts is conducive to enabling better application of viruses as biocontrol agents.Certain viruses carried by parasitic wasps enhance the parasitic efficiency of wasp-larvae by protecting them against the immune system of their Lepidopteran host.However,the relationship between prey pests and viruses found in predatory natural enemies remains unclear.Herein,we report the interaction between Arma chinensis virus-1(AcV-1),originally isolated from a predatory natural enemy,Arma chinensis(Hemiptera:Pentatomidae),and one of its prey species,Spodoptera frugiperda(Lepidoptera:Noctuidae).The results showed that the AcV-1 virus appeared harmful to the novel host S.frugiperda by inhibiting larval diet consumption and increasing pupal mortality.Meanwhile,sequencing data indicated that the virus altered the gene expression profiles of S.frugiperda.KEGG analysis showed that the proteasome and phagosome pathways related to protein degradation and immune response were significantly enriched.Although the expression levels of digestive enzyme genes did not change significantly,the total protease activity of AcV-1 virus-positive individuals was significantly decreased,suggesting that the virus inhibited diet consumption of S.frugiperda via the down-regulation of digestive enzyme activities.These results indicate that a virus initially isolated in a predatory natural enemy can decrease the fitness of its prey species.The virus was found to impact the host proteasome and phagosome pathways related to protein degradation and immunity,providing a potential mechanism to enhance controlling efficiency.

The Response of Carbonate System to Watershed Urbanization Process in a Semi-Arid River

Different from rivers in humid areas,the variability of riverine CO_(2) system in arid areas is heavily impacted by anthropogenic disturbance with the increasing urbanization and water withdrawals.In this study,the water chemistry and the controls of carbonate system in an urbanized river(the Fenhe River)on the semi-arid Loess Plateau were analyzed.The water chemistry of the river water showed that the high dissolved inorganic carbon(DIC)concentration(about 37 mg L^(-1))in the upstream with a karst land type was mainly sourced from carbonate weathering involved by H_(2)CO_(3) and H_(2)SO_(4),resulting in an oversaturated partial pressure of CO_(2)(pCO_(2))(about 800μatm).In comparison,damming resulted in the widespread appearance of non-free flowing river segments,and aquatic photosynthesis dominated the DIC and pCO_(2) spatiality demonstrated by the enriched stable isotope of DIC(δ^(13)CDIC).Especially in the mid-downstream flowing through major cities in warm and low-runoff August,some river segments even acted as an atmospheric CO_(2) sink.The noteworthy is wastewater input leading to a sudden increase in DIC(>55 mg L^(-1))and pCO_(2)(>4500μatm)in the downstream of Taiyuan City,and in cold November the increased DIC even extended to the outlet of the river.Our results highlight the effects of aquatic production induced by damming and urban sewage input on riverine CO_(2) system in semi-arid areas,and reducing sewage discharge may mitigate CO_(2) emission from the rivers.

Limonene’s Bacteriostatic Activity against Ralstonia solanacearum by Compromised Membrane Integrity: A Transcriptomic and Metabolomic Analysis

Ralstonia solanacearum, the causative agent of bacterial wilt, is a soil-borne pathogen that poses a widespread threat to plants in the Solanaceae family. To elucidate the mechanism by which limonene exerts its effects on R. solanacearum, we first assessed the impact of limonene on the physiological indicators of the pathogen and subsequently analyzed its transcriptome and metabolome. Our findings indicate that limonene has a potent inhibitory effect on R. solanacearum, and it also suppresses the formation of the bacterial community biofilm. Limonene primarily regulates the terpene biosynthesis pathway in R. solanacearum, thereby potentially affecting signal transduction in the pathogen and disrupting its normal growth and development. These results significantly enhance our understanding of limonene’s response to the induction of bacterial wilt and provide a reference for further prevention and control of R. solanacearum.

Identification of stably expressed QTL for resistance to black shank disease in tobacco(Nicotiana tabacum L.) line Beinhart 1000-1

Cigar line Beinhart 1000-1 has effective durable resistance to black shank(BS) and is considered one of the most resistant sources in tobacco(Nicotiana tabacum L.). To investigate the inheritance and identification of stable quantitative trait loci(QTL) for BS response, F2,BC1 F2 individuals and BC1 F2:3 lines were produced from a cross between Beinhart 1000-1 and Xiaohuangjin 1025. Two major quantitative trait loci(M-QTL) named qBS7 and qBS17 were repeatedly detected under different conditions. QTL qBS7 was mapped to the region between PT30174 and PT60621 and explained 17.40%–25.60% of the phenotypic variance under different conditions. The other QTL qBS17 in interval PT61564–PT61538 of linkage group 17 was detected in a BC1 F2 population in the field and in BC1 F2:3 in both the field and at the seedling stage, explaining 6.90% to 11.60% of the phenotypic variance. The results improve our understanding of the inheritance of resistance to BS and provide information that can be used in marker-assisted breeding.

Quality Characteristics of Tobacco Leaves with Different Aromatic Styles from Guizhou Province,China

The relationships between chemical components and quality indexes were studied in the tobacco leaves with different aromatic styles. A total of 16 chemical components, 4 quality indexes, and 6 smoking quality indexes from 366 tobacco leaf samples with 4 different types of aroma from Guizhou Province, China, were subjected to principal component analysis and stepwise regression analysis. The tobacco leaves with different types of aroma showed remarkable difference in the contents of chemical components, quality indexes, and smoking quality indexes. The first principal factors （carbohydrates and nitrogen-containing compounds） of the chemical composition of the leaf were similar among different types of aroma, which showed that the quality of the leaf was mainly influenced by carbohydrates and nitrogen-containing compounds and their ratios. The factors for the second through the seventh principal components varied largely among varioUs aromatic types, suggesting the contribution of other chemical components to the leaf quality. In addition, the smoking quality of four different aromatic leaves showed significant correlation with the different chemical components. The quality of tobacco leaves with different types of aroma was influenced by multiple factors, especially ecological conditions and culture techniques, which may provide guidance for directive cultivation of high-quality tobacco leaves.

High-density SNP genetic linkage map construction and quantitative trait locus mapping for resistance to cucumber mosaic virus in tobacco(Nicotiana tabacum L.)

Genetic linkage maps are essential for studies of genetics, genomic structure, and genomic evolution, and for mapping quantitative trait loci (QTL). Identification of molecular markers and construction of genetic linkage maps in tobacco (Nicotiana tabacum L.), a classical model plant and important economic crop, have remained limited. In the present study we identified a large number of single nucleotide polymorphism (SNP) markers and constructed a high-density SNP genetic map for tobacco using restriction site-associated DNA sequencing. In 1216.30 Gb of clean sequence obtained using the Illumina HiSeq 2000 sequencing platform, 99,647,735 SNPs were identified that differed between 203 sequenced plant genomes and the tobacco reference genome. Finally, 13,273 SNP markers were mapped on 24 high-density tobacco genetic linkage groups. The entire linkage map spanned 3421.80 cM, with a mean distance of 0.26 cM between adjacent markers. Compared with genetic linkage maps published previously, this version represents a considerable improvement in the number and density of markers. Seven QTL for resistance to cucumber mosaic virus (CMV) in tobacco were mapped to groups 5 and 8. This high-density genetic map is a promising tool for elucidation of the genetic bases of QTL and for molecular breeding in tobacco.

Changes of Polyphenols in Tobacco Leaves During the Flue-Curing Process and Correlation Analysis on Some Chemical Components

The changes of polyphenols in tobacco leaves during the flue-curing process and correlation analysis on some chemical components were studied. Leaf samples were taken from different tobacco-producing regions in Henan Province, China. The results indicated that the content of total phenols increased during the first 24 h of curing, and then decreased. It reached the lowest value at 72 h of curing and increased rapidly after that. The content of chlorogenic acid also increased during 0-24 h of curing. But the lowest point occurred at 60 or 72 h of curing and then it increased till the end of the curing process. The content of mtin generally increased with curing, and showed little fluctuations. The changes of PPO and POD activity were the opposite. Rutin was found to have a highly significant positive correlation with total sugar （r=0.822^＊＊）, but a highly significant negative correlation with starch, nicotine, and protein.

Comparative transcriptome analysis reveals differential gene expression in resistant and susceptible tobacco cultivars in response to infection by cucumber mosaic virus

Cucumber mosaic virus(CMV) is one of the most severe viral diseases transmitted by aphids infecting Solanum crops in China, causing great losses of crop yields and income in rural communities.The tobacco cultivars NC82 and Taiyan 8 are closely related but differ in resistance to CMV.NC82 is susceptible to infection and Taiyan 8 is resistant, but the mechanisms underlying this difference in resistance are not clear.In this study, we conducted RNA sequencing to analyze changes in gene expression induced in the leaves of Taiyan 8 and NC82 upon systemic infection with CMV, compared with gene expression in the leaves of mock-inoculated plants.Leaves were sampled at one, three, eight, and 15 days after infection.In total, 3443 and 747 differentially expressed genes were identified in Taiyan 8 and NC82, respectively.Gene ontology and pathway enrichment analyses revealed that the different responses to CMV infection between cultivars were based on microtubulebased processes, pentose and glucuronate interconversions, plant–pathogen interaction,and hormone signal transduction pathways.Genes encoding pathogenesis-related proteins, disease-resistance proteins, lipoxygenase, cellulose synthase, an auxin response factor, and an ethylene receptor showed different expression patterns.The differences in gene expression following CMV infection likely contributed to the different resistance levels of these two tobacco cultivars.The comprehensive transcriptome dataset described here,which includes candidate response genes, will serve as a resource for further studies of the molecular mechanisms associated with tobacco defense responses against CMV.

Screening of Antagonistic Bacteria from Phyllosphere towards Tobacco Brown Spot Fungus Alternaria alternata

The possibility of employing antagonistic bacteria for the control of tobacco brown spot was studied. Approximately 136 strains of bacteria were isolated from phyllospheres of tobacco and 9 of these possessed high levels of antagonistic properties, They significantly reduced brown spot in detached tobacco leaves when artificially inoculated with Alternaria alternata. Culture filtrate of the most effective bacterial isolate which designated as Tpb88 was shown to be very efficient in inhibiting mycelial growth of A alternata in dual cultures. Culture filtrate of Tpb88 inhibited germination and germ tube elongation of A alternata. The results showed that the culture filtrate directly inhibited spore germination of A alternata, especially during the first hours of the paired cultivation. The rate of antagonistic activity of culture filtrate of Tpb88 depended on its concentration in the mixture. The greatest inhibition of spore germination was ob served at the highest concentration of filtrate （filtrate to fungal spores inocula mixed in proportion 1 ： 50）. These suggest that the hypothetic mechanism of Tpb88 against tobacco brown spot is to produce antagonistic substances.

Effects of Different Flue-curing Technologies on Submicroscopic Structure of Flue-cured Tobacco Leaves

[ Objective] This study aimed to compare the effects of low-temperature low-humidity tobacco flue-curing technology, moderate-temperature moderate- humidity tobacco flue-curing technology and moderate-temperature high-humidity tobacco flue-curing technology on submicroscopic structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method] Middle leaves of tobacco cuhivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the submicroscopic structure of flue-cured tobacco leaves. [ Result] Morphology changes and structural rupture of chloroplasts occurred at different time during three flue-curing proces- ses. In low-temperature low-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred late but rapidly; in moderate-tempera- ture moderate-humidity flue-curing process and moderate-temperature high-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred early but slowly. Among these three flue-curing processes, the speed of changes in mitochondria morphology, cristae number and membrane rupture pres- ented an upward order of low-temperature low-humidity flue-curing process 〈 moderate-temperature high-humidity flue-curlng process 〈 moderate-temperature moderate-humidity flue-curing process. In low-temperature low-humldity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 36 -48 h ; in moderate-temperature moderate-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 24 - 36 h; in moderate-temperature high-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 12 - 24 h. [ Conclusion ] Using low-temperature low-humidity flue- curing technology could prolong the maintenance duration of the submicroscopic structure integrity of flue-cured tobacco leaves, which was conducive to fully decom- posing and transforming substances in tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

Effects of Different Flue-curing Technologies on Microscopic Structure of Flue-cured Tobacco Leaves

[Objective] This study aimed to compare the effects of Chinese quality-improving fragrunce-increasing tobacco flue-curing technology, Chinese three- stage tobaeeo flue-curing technology and Zimbabwean tobaceo flue-euring technology on microscopie structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method ] Middle leaves of tobacco cultivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the microscopic structure of flue-cured tobaceo leaves. [ Result] During three flue-curing processes, leaves, palisade tissues and sponge tissues shrank gradually. Three flue-euring processes exhibited significant differ- ences in the peak of tissue shrinkage ： microscopic structure of flue-cured tobacco leaves in Chinese three-stage tobacco flue-curing process shrank earliest, follower by Chinese quality-improving fragrance-increasing tobacco flue-curing process; flue-eured tobacco leaves in Zimbabwean tobacco flue-curing process presented the latest shrinkage. At 84 h post-curing, leaf thickness in three flue-curing processes showed a descend order of Chinese three-stage tobacco flue-curing process 〉 Chinese quality-improving fragrance-increasing tobacco flue-curing process 〉 Zimbabwean tobacco flue-curing process. Upper and lower epidermal cells in Zimb- abwean tobacco flue-curing process ruptured earlier than other two flue-curing processes; eventually, the majority of cells ruptured and mixed with palisade tissues and sponge tissues. In Chinese quality-improving fragranee-increasing tobacco flue-curing process, only a small number of epidermal cells in dried leaves ruptured. Among three flue-curing technologies, Zimbabwean tobacco flue-curing technology exhibited the greatest damage to epidermal cells, followed by Chinese three-stage tobaceo flue-curing technology; Chinese quality-improving fragrance-increasing tobacco flue-curing technology had the minimum damage to epidermal cells. Mese- phyll cross-section exhibited significant morphological changes in stomata. To be specific, at 0 - 12 h post-curing, stomata of tobacco leaves closed with slightly prominent stamatal apparatuses on upper and lower epidermis; at 24 -72 h post-curing, stomata of tobaceo leaves changed gradually from opening to closure with significantly prominent stomatal apparatuses on upper and lower epidermis; at 84 h post-curing, stomata of tobacco leaves closed, and the majority of stomatal appa- ratuses were significantly prominent. In Chinese quality-improving fragrance-increasing tobacco flue-curing process, only a small number of epidermal ceils ruptured at 84 h pest-flue-curing; palisade tissues and sponge tissues shrank almost simultaneously. Significant gaps were observed between palisade tissues and between pal- isade tissues and sponge tissues. Chinese quality-improving fragrance-increasing tobacco flue-curing technology exhibited lower disorder level compared with other two flue-curing technologies. [ Conclusion] Chinese quality-improving fragrance-increasing tobacco flue-curlng technology was conducive to maintaining the micro- scopic structure integrity of flue-cured tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

Recent Advances on Key Enzyme Genes in Metabolic Pathways Related to the Synthesis of Aroma Compounds in Tobacco

This article summarized three main kinds of metabolic pathways related to the synthesis of aroma compounds in plants, concluded the roles and expres- sion patterns of key enzyme genes catalyzing the formation of major intermediate products in phenylpropanoid metabolic pathway, isoprene metabolic pathway and alkaloid biosynthetic pathway respectively, highlighted the latest developments of these key enzyme genes in tobacco, and accordingly proposed that in-depth study at the protein level and analysis of metabolic network interaction should be carried out in tobacco besides the expression regulation and transgenic crop improvement at the genetic level. Based on the above analysis, further improvement of tobacco aroma quality through metabolic engineering and its application prospect in agricultural production were prospected.

Differential responses of root growth to nutrition with different ammonium/nitrate ratios involve auxin distribution in two tobacco cultivars

Nitrogen(N), the major forms of which are nitrate(NO3–) and ammonium(NH4+), plays an important role in plant growth and mediation of root development. However, the role of auxin in root growth in response to different NH4+/NO3– ratios remains unclear. Two tobacco cultivars(Nicotiana tabacum L.) were adopted in this study, which displayed variant growth features under the situations with sole NO3– nutrition ratio(NH4+/NO3– ratio: 0/100), low NO3– nutrition ratio(NH4+/NO3– ratio: 97/3), and optimal NH4+/NO3– ratio(50/50). We investigated the effects of the different NH4+/NO3– ratios on the formation and elongation of lateral roots(LRs), auxin concentration, DR5::GUS expression, 3 H-labeled indole acetic acid([3 H]IAA) transport, and the expression of six PIN genes in tobacco roots. We also examined the effects of exogenous auxin and a transport inhibitor on LRs growth. The results are shown as follows, compared to optimal N nutrition conditions, the biomass and nitrogen(N) accumulation were largely reduced by sole and low NO3– nutrition treatment in NC89, but no difference was observed in Zhongyan 100. In most cases, sole and low NO3– nutrition impaired the elongation and formation of firstorder lateral roots(1° LRs), only in NC89, thus reducing the root growth. IAA concentration and DR5::GUS expression levels decreased in roots when NC89 was subjected to sole and low NO3– nutrition media, suggesting that different NH4+/NO3– ratios affect the transport of auxin from leaves to roots. Results were similar following exogenous NAA application to low NO3– nutrition treated seedlings. Based on direct [3 H]IAA transport measurement, the transport of polar auxin from shoots to roots decreased due to low NO3– nutrition. PIN4 expression levels were markedly decreased in roots of NC89by sole and low NO3– nutrition, while they were unaffected in Zhongyan 100 roots. Overall, our findings suggest that LRs formation in tobacco seedlings is regulated by NH4+/NO3– ratios via modifying polar transport of auxin.

Characterization and Expression Analysis of Four Glycine-Rich RNA-Binding Proteins Involved in Osmotic Response in Tobacco (Nicotiana tabacum cv. Xanthi)

Plants have developed many signals and specific genes＇ regulations at both transcriptional and post-transcriptional levels in order to tolerate and adapt to various environmental stresses. RNA-binding proteins （RBPs） play crucial roles in the post- transcriptional regulation via mRNA splicing, polyadenylation, sequence editing, transport, mRNA stability, mRNA localization, and translation. In this paper, four cDNAs of glycine-rich RNA-binding proteins （GR-RBPs）, named NtRGP-la, -lb, -2, and -3, were isolated from Nicotiana tabacum by RT-PCR analysis, and special emphases were given to the sequences alignment, phylogenetic analysis and gene expression. Sequences alignment revealed minor difference of cDNA sequences, but no difference of deduced proteins between N. sylvestris and N. tabacum. Phylogenetic alignment revealed that four cDNAs in tobacco were clustered into two different groups. NtRGP-2 and -3 were evolutionarily closest to Arabidopsis GR-RBPs genes and related to animal GR-RBPs genes, while NtRGP-la and -lb were closest to Gramineae GR-RBPs genes. The expression analyses of these four NtRGPs in response to different abiotic stresses revealed the similar expression pattern. Moreover, the four NtRGPs, especially NtRGP-la and NtRGP-3, were strongly induced by stresses including water, wound, cold, and high temperature, weakly induced by PEG, drought and SA, while reduced by NaC1 and unaffected by ABA treatment. The fact that all of these abiotic stresses included in our experiments affected the water balance and resulted in osmotic stress on cellular level, suggests that NtRGPs in tobacco should be a family of crucial osmosis-related proteins, and may play a key role in signal transduction with ABA-independent pathway under abiotic stresses.