Diversity and varietal classification of Hibiscus syriacus L. with the heterogeneity within retrotransposon-like elements.
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Retrotransposons are present in multi-copy numbers that are integrated into plant genomes with considerable heterogeneous sequences within a single plant and between plant species, which allows the use of retrotransposons as additional sources of DNA polymorphism. A primer design for the sequence-tagged specific site and cleaved amplified polymorphic sequences (STS-CAPs) that are derived from retrotransposon-like sequences was developed for the molecular marker analysis in Hibiscus syriacus. This method was applied for the detection of sequence variations of intact retrotransposons that exist in plant genomes, which resulted in higher polymorphisms than in the amplified fragment length polymorphism (AFLP). Through STS-CAPs, specific fingerprinting data among H. syriacus varieties can be easily distinguished and generated with reproducible results. It could also be adapted to any species that possess multi-copy retrotransposons for varietal identification as well as the assessment of genetic relationships. (+info)
The p23 protein of hibiscus chlorotic ringspot virus is indispensable for host-specific replication.
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Hibiscus chlorotic ringspot virus (HCRSV) possesses a novel open reading frame (ORF) which encodes a putative 23-kDa protein (p23). We report here the in vivo detection of p23 and demonstrate its essential role in viral replication. The expression of p23 could be detected in protein extracts from transfected kenaf (Hibiscus cannabinus L.) protoplasts and in HCRSV-infected leaves. Further, direct immunoblotting of infected kenaf leaves also showed the presence of p23, and transient expression in onion and kenaf cells demonstrated that the protein is distributed throughout the cell. Site-directed mutagenesis showed that mutations introduced into the ORF of p23 abolished viral replication in kenaf protoplasts and plants but not in Chenopodium quinoa L. The loss of function of the p23 mutant M23/S33-1 could be complemented in trans upon the induced expression of p23 from an infiltrated construct bearing the ORF (pCam23). Altogether, these results demonstrate that p23 is a bona fide HCRSV protein that is expressed in vivo and suggest that p23 is indispensable for the host-specific replication of HCRSV. In addition, we show that p23 does not bind nucleic acids in vitro and does not act as a suppressor of posttranscriptional gene silencing in transgenic tobacco carrying a green fluorescent protein. (+info)
Covariation in the capsid protein of hibiscus chlorotic ringspot virus induced by serial passaging in a host that restricts movement leads to avirulence in its systemic host.
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Hibiscus chlorotic ringspot virus (HCRSV) from naturally infected Hibiscus rosa-sinensis L. loses virulence in its experimental systemic host Hibiscus cannabinus L. (kenaf) after serial passages in a local lesion host Chenopodium quinoa. Here we report the genetic changes responsible for the loss of virulence at the molecular level. A remarkable covariation of eight site-specific amino acids was found in the HCRSV capsid protein (CP) after serial passages in C. quinoa: Val(49)-->Ile, Ile(95)-->Val, Lys(270)-->Arg, Gly(272)-->Asp, Tyr(274)-->His, Ala(311)-->Asp, Asp(334)-->Ala, and Ala(335)-->Thr. Covariation of at least three of the eight amino acids, Val(49), Ile(95), and Lys(270), caused the virus to become avirulent in kenaf. Interestingly, the nature of the covariation was consistent and reproducible at each serial passage. These data indicate that the nonsynonymous substitutions of amino acids in the HCRSV CP after serial passages in C. quinoa are not likely to be random events but may be due to host-associated positive selection or accelerated genetic drift. The observed interdependence among the three amino acids leading to avirulence in kenaf may have implications for structural or functional relationships in this virus-host interaction. (+info)
Digestibility and dry matter intake of diets containing alfalfa and kenaf.
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Two experiments were conducted to determine the dietary value of pellets containing kenaf (Hibiscus cannabinus cv. 'Everglade 41') hay. Averaged across both experiments, kenaf pellets contained 82.6% kenaf hay, 16.6% liquid molasses, and 0.8% mineral oil. The chemical composition of the kenaf pellet was 12.6% crude protein (CP), 41.2% neutral detergent fiber (NDF), and 14.4% acid detergent fiber (ADF). In Exp. 1 (digestion and N balance trial), 18 lambs (body weight [BW] = 36.4 kg) were blocked by BW. Lambs were randomly assigned within a block to Diet 1 (59.5% corn and 40.5% alfalfa pellet), Diet 2 (59.7% corn, 28.4% alfalfa pellets, and 11.9% kenaf pellets), or Diet 3 (59.6% corn, 16.5% alfalfa pellets, and 23.9% kenaf pellets). Diets were formulated so that CP was the first-limiting nutrient. Each diet was limit-fed at 2.4% of BW. Replacing alfalfa pellets with kenaf pellets tended to decrease (P = 0.10) CP and ADF intakes, but increased (P = 0.01) DM digestibility. Diet had no effect (P = 0.33) on N balance. In Exp. 2 (dry matter [DM] intake trial), 32 lambs (BW = 30.4 kg) were blocked by gender and BW. Within a block, lambs were randomly assigned to one of four diets in a 2 x 2 factorial arrangement. Main effects were hay (bermudagrass or fescue) and supplemental protein source (kenaf or alfalfa pellets). Lambs were housed in individual pens with ad libitum access to the assigned hay. Supplemental protein was fed (185 g of DM) once daily. Hay intake was measured weekly for 8 wk. Lambs consumed more (P = 0.002) fescue than bermudagrass hay (743 vs 621 g/ d). Lambs fed fescue hay gained weight more rapidly (P = 0.001) than lambs fed bermudagrass hay (120 vs 72 g/d). Hay intake and ADG were similar (P = 0.90) for lambs fed alfalfa or kenaf pellets. Kenaf hay mixed with molasses and mineral oil can be formed into a pellet. In the diets used in this experiments, kenaf pellets can replace alfalfa pellets in diets fed to lambs without altering forage intake, gain, or N retention. (+info)
Synergism of the 3'-untranslated region and an internal ribosome entry site differentially enhances the translation of a plant virus coat protein.
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The use of internal ribosome entry sites (IRESs) is one of the unorthodox mechanisms exploited by viruses to initiate the translation of internal genes. Herein, we report a plant virus exploiting an IRES and its 3'-untranslated region (UTR) to express its internal genes, notably the 3'-proximal viral coat protein gene. Hibiscus chlorotic ringspot virus (HCRSV), a positive-strand non-polyadenylated RNA virus, was demonstrated to harbor a unique 100-nucleotide (nt) IRES, located 124 nt upstream of the coat protein gene, that could function in wheat germ extract, rabbit reticulocyte lysate, and mammalian cells. In comparison with other known IRESs of picornaviruses and eukaryotic mRNAs, this 100-nt IRES is distinctively short and simple. The IRES activity was tested in homologous and heterologous bicistronic constructs, and the expression of the 3'-proximal gene was enhanced when the 3'-UTR was present. When the IRES element was bisected, each half still possessed IRES activity and could initiate internal translation on its own. Site-directed mutagenesis and deletion analyses revealed that the primary sequence within the 5' half was crucial for IRES activity, whereas the primary sequence of the second half and a GNRA motif were non-essential. To our knowledge, this is the first report describing a mechanism whereby an IRES, located in the 3' portion of the virus genome, co-operates with the 3'-UTR to enhance gene expression differentially. (+info)
Effect of maturity on macromineral content of selected leafy vegetables.
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Macro mineral contents were estimated in commonly consumed green leafy vegetables in India, namely; Koyyathotakura and Peddathotakura (varieties of Amaranthus species); Erragogu and Tellagogu (variety of Hibiscus species); Gangabayalakura (Portulaca olereceo) and Palak (Spineces olerecea) at three different stages of maturity. Varietal differences were also observed. The results of the study showed that as the plant matured from stage I (15 days) to stage II (30 days) calcium and magnesium content increased. In contrast, phosphorus content decreased as the plant matured. Varietal differences were also observed at different stages of maturity. The results also indicated that the consumption of green leafy vegetables at stage I (15 days) and stage II (30 days) potentially provide the greatest amount of minerals. (+info)
Genetic diversity of Hibiscus tiliaceus (Malvaceae) in China assessed using AFLP markers.
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Amplified fragment length polymorphism (AFLP) markers were used to investigate the genetic variations within and among nine natural populations of Hibiscus tiliaceus in China. DNA from 145 individuals was amplified with eight primer pairs. No polymorphisms were found among the 20 samples of a marginal population of recent origin probably due to a founder effect. Across the other 125 individuals, 501 of 566 bands (88.5%) were polymorphic, and 125 unique AFLP phenotypes were observed. Estimates of genetic diversity agreed with life history traits of H. tiliaceus and geographical distribution. AMOVA analysis revealed that most genetic diversity resided within populations (84.8%), which corresponded to results reported for outcrossing plants. The indirect estimate of gene flow based on phiST was moderate (Nm=1.395). Long-distance dispersal of floating seeds and local environments may play an important role in shaping the genetic diversity of the population and the genetic structure of this species. (+info)
Paralogy and orthology in the MALVACEAE rpb2 gene family: investigation of gene duplication in hibiscus.
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A sample of the second largest subunit of low-copy nuclear RNA polymerase II (rpb2) sequences from Malvaceae subfamily Malvoideae suggests that rpb2 has been duplicated early in the subfamily's history. Hibiscus and related taxa possess two rpb2 genes, both of which produce congruent phylogenetic patterns that are largely concordant with cpDNA topologies. No evidence of functional divergence or disruption was found among duplicated copies, suggesting that long-term maintenance of duplicated copies of rpb2 is usual in this lineage. Therefore, this gene may be suitable for the potential diagnosis of relatively old polyploid events. One probable pseudogene was found in Radyera farragei and a single chimeric sequence was recovered from Howittia trilocularis, suggesting that the rpb2 locus is not as prone to evolutionary processes that can confound phylogenetic inferences based on nDNA sequences. The pattern of relationships among rpb2 sequences, coupled with chromosome number information and Southern hybridization data, suggests that an early polyploid event was not the cause of the duplication, despite independent evidence of paleopolyploidy in some members of Malvoideae. Rpb2 exons and introns together are suitable for phylogenetic analysis, producing well-resolved and well-supported results that were robust to model permutation and congruent with previous studies of subfamily Malvoideae using cpDNA characters. (+info)