The Effects of Nanosilver on Bacterial Contamination and Increase Durability Cultivars of Rosa hybrida L. Through of Stenting Method.

The rose flower (Rosa hybrida L.) is one of the world's most popular flowers among ornamental plants and the cut rose flower industry is the most important aspect of the rose culture industry in the world. The stenting method (cutting and grafting) uses a stem containing a leaf and a dormant bud as a scion, which is grafted onto an internode from the rootless rootstock so that the formation of the graft and the induction of the root on the rootstock are carried out simultaneously. In this method, Natal Briar is often used due to its desirable traits such as high rooting, the possibility of faster regeneration after cutting, the ability to adapt to different culture media and high resistance to root diseases. Most imported Natal Briar rootstocks, in addition to increasing the cost of producing hybrid roses, have a percentage of Agrobacterium contamination that can reduce the efficiency of stenting. Therefore, using the tissue culture technique to propagate the base is one of the important solutions. But one of the problems of tissue culture is the control of bacterial contamination so that these factors grow faster than explants and absorbing carbohydrate sources from the culture medium prevents regeneration of Natal Briar rootstocks. Nanotechnology today has expanded the field of application of nanomaterials due to the increase in the surface-to-volume ratio, electron transfer ability and surface reaction capability. Inhibit DNA replication and destroy the cellular structure of Agrobacterium. Therefore, the use of appropriate concentrations of Nanosilver in the culture medium of Natal Briar rootstocks increases the economic efficiency and durability of Rosa hybrida cultivars obtained by the stenting method.

Somatic embryogenesis and ß-glucuronidase transformation in chickpea (Cicer arietinum cv. Bivanich).

BACKGROUND: Chickpea (Cicer arietinum L.) is an important kernel legume in the world. To optimize the plant tissue culture some experiments such as direct regeneration, proliferation, rooting shoots and somatic embryogenesis were done. METHODS AND RESULTS: In experiments were used direct regeneration and proliferation, various levels of plant growth regulators NAA (0 and 1.0 mg/l), BAP (0, 1, 3 and 5 mg/l) and three explants' types (epicotyl, cotyledon and embryonic axis). The results of both experiments showed that embryonic axis explant was better than other explants. The highest percentage was obtained in MS media containing 1 mg/l BAP and also 3 mg/l BAP and 0.1 mg/l NAA with an average of 72%. The highest average number of branches (4.66) was found in the proliferation of embryonic axis in MS medium containing 3 mg/l BAP. The highest rooting shoot (90%) was found in 1/2MS in B5 medium vitamins with 0.2 mg/l of IBA and 0.5 mg/l NAA. Somatic embryogenesis experiments were compared on the concentration gradient of 2,4-D in fine embryonic axis explants. The results displayed that the concentration gradient of 10 mg/l 2,4-D to 5 mg/l of 2,4-D and then to zero concentration showed the highest number of embryos. CONCLUSION: The best environment for regeneration embryos was MS medium with 2.5 mg/l of 2,4-D concentration gradient to zero. In this study, the PCR reaction showed the presence of the ß-glucuronidase (gus) marker gene in regenerated cotyledons for 20 min in all three strains studied.

Endoplasmic reticulum retention signaling and transmembrane channel proteins predicted for oilseed ω3 fatty acid desaturase 3 (FAD3) genes.

Oilseed crop oils contain a variety of unsaturated fatty acids that are synthesized and regulated by fatty acid desaturases (FADs). In this study, 14 FAD3 (ω3 desaturase) protein sequences from oilseeds are analyzed and presented through the application of several computational tools. The results indicated a close relationship between Brassica napus and Camelina sativa, as well as between Salvia hispanica and Perilla frutescens FAD3s, due to a high similarity in codon preferences in codon usage clusters and the phylogenetic tree. The cis-acting element results reveal that the seed-specific promoter region of BnFAD3 contains the critical conserved boxes such as HSE and ABRE, which are involved in responsiveness to heat stress and abscisic acid. The presence of the aforementioned conserved boxes may increase cold acclimation as well as tolerance to drought and high salinity. Omega(ω)3 desaturases contain a Skn-1 motif which is a cis-acting regulatory element required involved in endosperm development. In oilseed FAD3s, leucine is the most repeated amino acid in FAD3 proteins. The study conveyed that B. napus, Camelina sativa, Linum usitatissimum, Vernicia fordii, Gossypium hirsutum, S. hispanica, Cannabis sativa, and P. frutescens have retention signal KXKXX/XKXX at their c-terminus sites, which is one of the most important characteristics of FADs. Additionally, it was found that BnFAD3 is a transmembrane protein that can convert ω6 to ω3 fatty acids and may simultaneously act as a potassium ion channel in the ER.

The study of inter-specific relationships of Bromus genus based on SCoT and ISSR molecular markers.

The genus of Bromus is one of the most important collection of rangeland plants, which are distributed in a wide range of natural areas of Iran. Interspecific relationships were evaluated in 90 accessions of 18 Bromus species based on 15 ISSR and 15 SCoT primers. SCoT markers separated the accessions better than ISSR marker. In addition, there was a high interspecific diversity between surveying germplasm. The sections of Bromus genus completely separated based on DNA molecular markers. SCoT markers could separate the accessions in each species. The primers of SC5 and SC35 from SCoT marker and UBC861, UBC857 and UBC844 primers from ISSR marker were identified as the best primers in revealing of genetic diversity between accessions. The sections of Ceratochloa, Genea, Pnigma and Bromus were monophyletic and were placed in one cluster. The section Bromus had a direct relationship with section Genea. In other words, section Ceratochloa has a direct relationship with Pnigma. B. tectorum and B. sericeus. B. sterilis had the most distance with other species in section Genea. B. squarrosus and B. japonicus had the most similarity and B. briziformis with B. danthoniae and B. scoparius with B. rechingeri had a moderate relationship in section Bromus. B. tomentosus and B. persicus had the highest similarity and B. riparius with B. biebersteinii and B. tomentellus with B. inermis had a moderate similarity in section Pnigma.

Influence of abiotic elicitors on improvement production of artemisinin in cell culture of Artemisia annua L.

A significant sesquiterpene lactone used as a drug is artemisinin. It is definitely an anti-parasitic drug isolated from field-grown Artemisia annua L. a plant from Asteraceae family. It is the best treatment for Plasmodium falciparum malaria. Unfortunately, artemisinin content in A. annua is extremely low (0.01-0.8% dry weight). So, some researchers focused on enhancing artemisinin content either in tissue/cell culture or the whole plant of A. annua sp. The aims of the current study were the effect of plant growth regulators on callus production and improvement of artemisinin content in cell suspension culture of A. annua, an alternative to the whole plant using abiotic elicitors. For callus induction, an experiment was laid out as a factorial experiment with three factors (explant type, different concentrations of BAP and 2,4-D) based on completely randomized design with three replications. The maximum frequency of callus induction (100%) was found in leaf explant on MS medium with a combination of 2, 4-D (3 mg/l) and BAP (1.5 mg/l). Therefore, the best calli were used for cell suspension culture and the effects of GA3 and ABA as abiotic elicitors were evaluated on the improvement of artemisinin production. The results indicated that both ABA and GA3 increased artemisinin content (2.02 fold and 1.67 fold in comparison to control respectively) in cell suspension culture.

Cell dedifferentiation and multiplication of Burdock (Arctium Lappa) as a medicinal plant.

Arctium lappa L. (Burdock) is an important plant with various pharmacological effects. According to the importance of this plant, optimization of its tissue culture will lead to more investigation and application of it. The aim of this study was to develop protocols for callus induction and shoot regeneration of A.  lappa. In order to optimize of tissue culture in A. lappa, callus induction, indirect regeneration and direct regeneration were carried out in factorial experiment based on Completely Randomized Designs (CRDs). Hypocotyl and cotyledon were cultured on the Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 6-Benzylaminopurine (BAP) for callus induction. In indirect regeneration experiment various levels of BAP and α-Naphthaleneacetic acid (NAA) and two types of explants (calli derived from cotyledon and hypocotyl) were investigated. In direct regeneration section, various levels of BAP plus 2 mg/l NAA and different explants (cotyledon, hypocotyl and bud) were compared. In both cotyledon and hypocotyl, the maximum callus induction was observed on a media containing 2 mg/l 2,4-D plus 1 mg/l BAP (100% and 76.19% respectively). The highest percentage of indirect regeneration (65%) was observed at 1 mg/l BAP plus 0.5 mg/l NAA on calli from hypocotyl. The highest percentage of direct regeneration (90.33) was observed in hypocotyl with a lateral bud explant on MS medium supplemented with 0.5 mg/l BAP plus 2 mg/l NAA. In this study, optimization of tissue culture protocol for A. lappa was carried out as a research technique, as well as technique for further exploitation of this plant.

Callus induction and somatic embryogenesis in Stevia rebaudiana Bertoni as a medicinal plant.

Stevia rebaudiana (Bert.) from Asteraceae family is a useful medicinal plant that prevents and cures diabetes, blood pressure, weight gain and tooth decay. Due to self-incompatibility in stevia, somatic embryo investigation for artificial seed production is valuable in this plant. In order to evaluate the callus induction characteristics in stevia, a factorial experiment was laid out based on a completely randomized design with three replications. The factors included ten hormone combinations and control, two kinds of media (MS and B5) and two types of explants (leaf and internode). Callus induction characters including the percentage of callus formation, days to callus induction, fresh and dry callus weight were recorded. Analysis of variance showed significant differences (p<0.01) among hormone combinations, media and explant types as well as their interactions. The best treatment for callus induction with minimum time to callus formation was 1 mg/l NAA+1 mg/l BAP. The highest fresh and dry callus weight were obtained on B5 medium supplemented by 1 mg/l 2,4-D+1 mg/l BAP (in leaf explant) and 0.25 mg/l 2,4-D+ 0.1 mg/l BAP (in internode explant). These results can be used in suspension culture. To induce somatic embryogenesis in suspension culture, six hormone treatments were investigated. The highest somatic embryogenesis percentage was obtained in MS medium supplemented by 2 mg/l 2,4-D+ 0.5 mg/l NAA+0.5 mg/l BAP.

Effect of waterlogging at different growth stages on some morphological traits of wheat varieties.

Excess rainfalls may be the cause of waterlogging in soil, which affects the growth and development of wheat. Therefore, the objectives of this study were to examine the effects of waterlogging on shoot and root growth and physiological characteristics of wheat. Three experiments were conducted: experiment 1 (E1): evaluation of seedling growth on ten Iranian winter wheat varieties with waterlogging periods (1-4, 4-8, 8-12, and 12-16 days starting from seed germination). Seminal roots and plumule were investigated at seedling. The others are E2: pretreatment of waterlogging (15 days) at tillering and stem elongation stages and its effects on shoot and root growth at anthesis stage and experiment 3 (E3): pretreatment of waterlogging (15 days) at tillering and jointing stages and its effects on yield and yield components and also evaluation of stress tolerance indexes. The results of the seedling growth test (E1) showed that 1-4- and 4-8-day waterlogging severity reduced seminal root length (94.5 to 93.7 %) and plumule length (86.2 to 50.0 %) compared to control. Results of E2 indicated that waterlogging stress decreased shoot dry weight, root dry weight, total secondary root length, and chlorophyll a + b content of flag leaf by 28-31, 44-35, 20-31, and 28-35 %, respectively. Also, result of E3 showed that the grain yields of wheat varieties at two conditions of stress were different in base tolerance indexes. In general, the responses of wheat varieties to waterlogging were different at the three experiments. The varieties that had the most of dry weight and length of the root were tolerant. Thus, it is possible to use these characteristics as an index for selecting the varieties with tolerance to waterlogging.

Isolation and sequence analysis of napin seed specific promoter from Iranian Rapeseed (Brassica napus L.).

Rapeseed (Brassica napus L.) has become an important crop during the last 30years. In addition to a high lipid level, the seeds also have a significant protein content, which constitutes 20-25% of the dry seed weight. The synthesis of storage proteins is primarily controlled at transcriptional level and seed-specific expression has been shown to be conferred upon the promoter regions of many storage protein genes. Napin is one of the main storage proteins in rapeseed(')s embryo that is produced in seed developing stage. Its promoter region located at 5' upstream of the napin gene has already been isolated (GenBank number, EU416279.1). In current research, seed-specific promoter (napin) of Iranian B. napus L. was isolated from the genomic DNA and cloned into pBI121 plant binary vector to use in future researches. For this purpose, the napin promoter was amplified by PCR method using specific primers, cloned in pSK(+) vector and sequenced. Sequencing analysis showed that the cloned promoter contained all of conserved motifs such as TATA box (TATAAA), RY repeats (CATGCA), dist-B (TCAAACACC) and prox-B elements (GCCACTTGTC), G-box (CACGTG) and CAAT Motifs, which constituted the seed-specific promoter activity and according to this analysis, the seed-specific promoter activity of cloned sequence was predicted. Based on sequence distances of nucleotide sequences, our sequence had the highest similarity (99.8%) whit B. napus sequence (with EU416279.1 accession number). Finally the promoter obtained might be interesting not only as a useful tool for biotechnological application but also for fundamental research.

Isolation, cloning, and characterization of a partial novel aro A gene in common reed (Phragmites australis).

CONTEXT: Among the essential amino acids, phenylalanine, tryptophan, and tyrosine are aromatic amino acids which are synthesized by the shikimate pathway in plants and bacteria. Herbicide glyphosate can inhibit the biosynthesis of these amino acids. So, identification of the gene tolerant to glyphosate is very important. It has been shown that the common reed or Phragmites australis Cav. (Poaceae) is relatively tolerant to glyphosate. OBJECTIVE: The aim of the current research is identification, cloning, sequencing, and registering of partial aro A gene of the common reed P. australis. MATERIALS AND METHODS: The partial aro A gene of common reed (P. australis) was cloned in Escherichia coli and the amino acid sequence was identified/determined for the first time. RESULTS: This is the first report for isolation, cloning, and sequencing of a part of aro A gene from the common reed. A 670 bp fragment including two introns (86 bp and 289 bp) was obtained. The open reading frame (ORF) region in part of gene was encoded for 98 amino acids. Alignment showed high similarity among this region with Zea mays (L.) (Poaceae) (94.6%), Eleusine indica L. Gaertn (Poaceae) (94.2%), and Zoysia japonica Steud. (Poaceae) (94.2%). The alignment of amino acid sequence of the investigated part of the gene showed a homology with aro A from several other plants. This conserved region forms the enzyme active site. CONCLUSION: The alignment results of nucleotide and amino acid residues with related sequences showed that there are some differences among them. The relative glyphosate tolerance in the common reed may be related to these differences.

The influence of Agrobacterium rhizogenes on induction of hairy roots and ß-carboline alkaloids production in Tribulus terrestris L.

We have developed an efficient transformation system for Tribulus terrestris L., an important medicinal plant, using Agrobacterium rhizogenes strains AR15834 and GMI9534 to generate hairy roots. Hairy roots were formed directly from the cut edges of leaf explants 10-14 days after inoculation with the Agrobacterium with highest frequency transformation being 49 %, which was achieved using Agrobacterium rhizogenes AR15834 on hormone-free MS medium after 28 days inoculation. PCR analysis showed that rolB genes of Ri plasmid of A. rhizogenes were integrated and expressed into the genome of transformed hairy roots. Isolated transgenic hairy roots grew rapidly on MS medium supplemented with indole-3-butyric acid. They showed characteristics of transformed roots such as fast growth and high lateral branching in comparison with untransformed roots. Isolated control and transgenic hairy roots grown in liquid medium containing IBA were analyzed to detect ß-carboline alkaloids by High Performance Thin Layer Chromatograghy (HPTLC). Harmine content was estimated to be 1.7 µg g(-1) of the dried weight of transgenic hairy root cultures at the end of 50 days of culturing. The transformed roots induced by AR15834 strain, spontaneously, dedifferentiated as callus on MS medium without hormone. Optimum callus induction and shoot regeneration of transformed roots in vitro was achieved on MS medium containing 0.4 mg L(-1) naphthaleneacetic acid and 2 mg L(-1) 6-benzylaminopurine (BAP) after 50 days. The main objective of this investigation was to establish hairy roots in this plant by using A. rhizogenes to synthesize secondary products at levels comparable to the wild-type roots.

Haplotype analysis of QTLs attributed to salinity tolerance in wheat (Triticum aestivum).

A diverse collection of wheat germplasm, consisting of 100 bread wheat lines with varying levels of salinity tolerance were evaluated based on incomplete block design (lattice) with two replications in field conditions. Plant material was screened for salinity tolerance under normal and saline field conditions. Subsequently in order to assess the haplotype diversity of QTLs attributed to salinity tolerance in wheat (Triticum aestivum), a collection of 30 extremes tolerant and sensitive genotypes among them were selected for genotyping on the basis of morphological, physiological and phenological traits. Genotyping was done using microsatellite markers which had been detected as the flanking regions of large effect QTLs attributed to salinity tolerance on chromosomes 2A, 4D and 3B. Combined analysis of saline and normal conditions revealed that genotypes showed highly significant responses. Association analysis of SSR markers with traits, showed markers Xcfa2121b, Xgwm10 and Xgwm296 on chromosome 2A and markers Xgwm194 and xgwm624 for chromosome 4D, had significant association with most of measured traits. Haplotype diversity analysis showed markers Xgwm10, Xgwm445, Xbarc353.2, Xgwm312, Xgwm515 and Xwmc296 on chromosome 2A as well as markers Xwmc326 and Xgwm345, Xbarc48.4 on chromosomes 3B and 4D were identified as the best markers attributed to salinity tolerance and they can be informative markers for improvement of salinity tolerance through marker-assisted selection programs.

Enhanced ß-ionone emission in Arabidopsis over-expressing AtCCD1 reduces feeding damage in vivo by the crucifer flea beetle.

Plant carotenoid derived ß-ionone has been shown to have diverse biological effects on some insect herbivores and herbivore parasitoids. In this study, Arabidopsis transgenic plants over-expressing a carotenoid cleavage dioxygenase1 gene (AtCCD1) were generated to test whether ß-ionone emissions could be enhanced and used to control feeding by the crucifer flea beetle (Phyllotreta cruciferae Goeze). The transgenic plants exhibited a morphological phenotype indistinguishable from the wild type (WT) control over their complete life cycle. Gas chromatography and mass spectrometry analyses of headspace volatiles collected from 6-wk-old intact flowering plants revealed substantially enhanced ß-ionone emission from transgenic plants compared with WT, but no ß-ionone enhancement occurred at a young vegetative stage (4-wk-old seedlings). Bioassays in an enclosed environment showed that AtCCD1 over-expression resulted in ≍50% less leaf area damage by flea beetles on transgenic plants compared with WT plants. The mean number of damaged transgenic leaves per plant also was significantly lower in transgenic plants (P<0.05). Our results indicate that AtCCD1 over-expression and induced ß-ionone emission might find application in the control of pests for Brassica crops grown in greenhouse operations. Potentially, ß-ionone also could be used on crops grown in open-air ecosystems if this allomone is released in sufficient quantities to discourage herbivore foragers.

RNAi-mediated suppression of DET1 alters the levels of carotenoids and sinapate esters in seeds of Brassica napus.

Carotenoids and sinapate esters in Brassica napus affect the nutritional value of the seed. In this study, the B. napus regulatory gene DE-ETIOLATED1 (DET1), which is a negative regulator of light-mediated responses in plants and affects carotenoid and flavonoid pathways in tomato, was suppressed both constitutively and in a seed-specific manner by RNAi. Constitutive silencing of DET1 resulted in transgenic seeds with substantially elevated levels of lutein, beta-carotene, and zeaxanthin relative to nontransgenic seeds. Levels of these carotenoids were also enhanced but to a lesser extent in seeds of transgenic plants with seed-specific silencing of DET1. Moreover, sinapate esters 1,2-disinapoylgentiobiose and 1,2-di-O-sinapoylglucose were identified in the seeds using 1D and 2D NMR, as well as ESI-MS spectrum analyses. The levels of 1,2-di-O-sinapoylglucose in seeds in both sets of transgenic plants were lower compared to nontransgenic seeds. The results revealed that DET1 suppression in B. napus can increase the levels of carotenoids and reduce the levels of sinapate esters simultaneously in the seeds, thus enhancing their overall nutritional value.