Investigation of the Potential Health Benefits as Lipase Inhibitor and Antioxidant of <Emphasis Type="Italic">Leopoldia comosa</Emphasis> (L.) Parl.: Variability of Chemical Composition of Wild and Cultivated Bulbs

There is a great interest in the nutritional value of vegetables and fruits and how the habitat affects nutritive and biological properties. In vitro studies here reported were performed to evaluate the inhibitory activity of formulations from edible plant on pancreatic lipase. The aim of this study was also to evaluate the biovariability of L. comosa (L.) Parl. bulbs from Italy. The wild bulbs were compared with the same cultivated species that are commonly commercialized to identify samples with the best quality for a potential therapeutic application. Hydroalcoholic extract and polar fraction of wild bulbs showed a very important pancreatic lipase inhibitory activity, with IC₅₀ values of 0.166 ± 0.005 and 0.153 ± 0.005 mg/mL, respectively. In order to characterize the extracts, gas chromatography associated with mass spectrometry (GC/MS) analysis was performed, revealing the predominance of palmitic acid. Phenolic and flavonoid composition was also evaluated. L. comosa extract obtained from wild bulbs demonstrated both antioxidant and anti-obesity activities that might be attributed to a wide range of present phenolic compounds.     

Amplification of the <Emphasis Type="Italic">Phytophthora infestans</Emphasis> RG57 loci Facilitates <Emphasis Type="Italic">in planta</Emphasis> T-RFLP Identification of Late Blight Genotypes

Late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is a devastating disease in potato and tomato and causes yield and quality losses worldwide. The disease first emerged in central America and has since spread in North America including the United States and Canada. Several new genotypes of P. infestans have recently emerged, including US-22, US-23 and US-24. Due to significant economic and environmental impacts, there has been an increasing interest in the rapid identification of P. infestans genotypes. In addition to providing details regarding the various phenotypic characteristics such as fungicide resistance, host preference, and pathogenicity associated with various P. infestans genotypes, information related to pathogen movement and potential recombination may also be determined from the genetic analyses. Restriction fragment length polymorphism (RFLP) analysis with the RG57 loci is one of the most reliable procedures used to genotype P. infestans. However, the RFLP procedure requires propagation and isolation of the pathogen and relatively large amounts of DNA. Isolation of the late blight pathogen is sometimes impossible due to the poor condition of the infected tissues or the presence of fungicide residues. In this study, we describe a procedure to identify P. infestans at the molecular level in planta using terminal restriction fragment length polymorphism (T-RFLP) of the RG57 loci. This T-RFLP assay is sufficiently sensitive to detect and differentiate P. infestans genotypes directly in planta without propagation and isolation of the pathogen, to facilitate the timely implementation of best management practices.     

Effect of water management on soil respiration and <Emphasis Type="Italic">NEE</Emphasis> of paddy fields in Southeast China

Water management is an important factor in regulating soil respiration and the net ecosystem exchange of CO₂ (NEE) between croplands and atmosphere. However, how water management affects soil respiration and the NEE of paddy fields remains unexplored. Thus, a 2-year field experiment was carried out to study the effects of controlled irrigation (CI) during the rice season on the variation of soil respiration and NEE, with flooding irrigation (FI) as the control. A decrease of irrigation water input by 46.39% did not significantly affect rice yield but significantly increased irrigation water use efficiency by 0.99 kg m^?3. The soil respiration rate of CI paddy fields was larger than that of FI paddy fields except during the ripening stage. Natural drying management during the ripening stage resulted in a significant increase of the soil respiration rate of the FI paddy fields. Variations of NEE with different water managements were opposite to soil respiration rates during the whole rice growth stages. Total CO₂ emission of CI paddy fields through soil respiration (total R _soil) increased by 11.66% compared with FI paddy fields. The increase of total R _soil resulted in the significant decrease of total net CO₂ absorption of CI paddy fields by 11.57% compared with FI paddy fields (p < 0.05). There were inter-annual differences of soil respiration and the NEE of paddy fields. Frequent alternate wetting and drying processes in the CI paddy fields were the main factors influencing soil respiration and NEE. CI management slightly enhanced the rice dry matter amount but accelerated the consumption and decomposition of soil organic carbon and significantly increased soil respiration, which led to the decrease of net CO₂ absorption. CI management and organic carbon input technologies should be combined in applications to achieve sustainable use of water and soil resources in paddy fields.     

Hypolipidemic Activity of <Emphasis Type="Italic">Camellia euphlebia</Emphasis> Flower Extract in High–fat-fed Mice

Camellia euphlebia (family, Theaceae) has been used for the prevention and treatment of cardiovascular diseases in Southern China. However, there has been no report on the hypolipidemic activity of Camellia euphlebia flower. This study evaluated the hypolipidemic activity of different preparation of Camellia euphlebia flower extracts using in vivo models. Mice intragastrically administered aqueous extract at 400 mg/kg dose or ethanol extract at 100 and 400 mg/kg doses of Camellia euphlebia flower for 28 days exhibited significant decreases in the levels of total cholesterol, triglycerides and low-density lipoprotein cholesterol, while displaying increased level of high-density lipoprotein cholesterol in the serum. The Camellia euphlebia flower extracts also improved the antioxidant ability of hyperlipidemic mice as well as protecting the animals against liver damage by lowering the level of glutamic-pyruvic transaminase activity. Furthermore, 400 mg/kg ethanol extract effectively down-regulated the mRNA levels of fatty acid synthase, 3-hydroxy-3-methylglutaryl CoA reductase and glycerol-3-phosphate acyl transferase, suggesting that Camellia euphlebia flower extract may potentially inhibit lipid accumulation in the liver by regulating the expression of fatty acid synthase, 3-hydroxy-3-methylglutaryl CoA reductase and glycerol-3-phosphate acyl transferase. These results provided support for the potential hypolipidemic activity of Camellia euphlebia flower and could partly explain the basis of using Camellia euphlebia for the treatment of hyperlipidemia.     

Ephemeral-habitat colonization and neotropical species richness of <Emphasis Type="Italic">Caenorhabditis</Emphasis> nematodes

Background

The drivers of species co-existence in local communities are especially enigmatic for assemblages of morphologically cryptic species. Here we characterize the colonization dynamics and abundance of nine species of Caenorhabditis nematodes in neotropical French Guiana, the most speciose known assemblage of this genus, with resource use overlap and notoriously similar external morphology despite deep genomic divergence.

Methods

To characterize the dynamics and specificity of colonization and exploitation of ephemeral resource patches, we conducted manipulative field experiments and the largest sampling effort to date for Caenorhabditis outside of Europe. This effort provides the first in-depth quantitative analysis of substrate specificity for Caenorhabditis in natural, unperturbed habitats.

Results

We amassed a total of 626 strain isolates from nine species of Caenorhabditis among 2865 substrate samples. With the two new species described here (C. astrocarya and C. dolens), we estimate that our sampling procedures will discover few additional species of these microbivorous animals in this tropical rainforest system. We demonstrate experimentally that the two most prevalent species (C. nouraguensis and C. tropicalis) rapidly colonize fresh resource patches, whereas at least one rarer species shows specialist micro-habitat fidelity.

Conclusion

Despite the potential to colonize rapidly, these ephemeral patchy resources of rotting fruits and flowers are likely to often remain uncolonized by Caenorhabditis prior to their complete decay, implying dispersal-limited resource exploitation. We hypothesize that a combination of rapid colonization, high ephemerality of resource patches, and species heterogeneity in degree of specialization on micro-habitats and life histories enables a dynamic co-existence of so many morphologically cryptic species of Caenorhabditis.

     

Dyeing and antibacterial properties of <Emphasis Type="Italic">Liriope platyphylla</Emphasis> fruit extracts on silk fabrics

Dyeing and antibacterial properties of a natural dye extracted from Liriope platyphylla fruit applied on silk fabrics have been studied. The total phenolic content (1109.13±69.02 mg), total flavonoid content (530.60±89.44 mg), and total anthocyanin content (492.26±77.79 mg) were measured in 100 g fresh weight of L. platyphylla fruits. In addition, ten anthocyanins and four flavanols were identified in L. platyphylla fruits by high performance liquid chromatography with diode array detection coupled with electrospray ionization mass spectrometry (HPLC-DAD/ESI-MS). A broad variation in color shade and color depth can be achieved with mixtures of dye extracts and metal mordants. Purple, blue, and pale green were main color shades of silk fabrics dyed with the extracts. The fastness of dyed silk fabrics except for control dyed fabrics against light, washing, and rubbing were acceptable with at least a grey scale rating of 3. The antibacterial activities of L. platyphylla fruit extracts were retained on dyed silk fabrics even after home washing 30 cycles. Mordanting with metal salt mordant had a positive effect on antibacterial activity of dyed silk fabrics in this study. Among them, aluminum and copper were the most effective mordants for improving antibacterial activity of silk fabrics dyed with L. platyphylla fruit extracts. The costs of natural dyeing of per silk fabrics kg by the extracts from L. platyphylla fruit were also calculated on laboratory scale.     

<Emphasis Type="Italic">GNS4</Emphasis>, a novel allele of <Emphasis Type="Italic">DWARF11</Emphasis>, regulates grain number and grain size in a high-yield rice variety

Background

Rice plays an extremely important role in food safety because it feeds more than half of the world’s population. Rice grain yield depends on biomass and the harvest index. An important strategy to break through the rice grain yield ceiling is to increase the biological yield. Therefore, genes associated with organ size are important targets for rice breeding.

Results

We characterized a rice mutant gns4 (grain number and size on chromosome 4) with reduced organ size, fewer grains per panicle, and smaller grains compared with those of WT. Map-based cloning indicated that the GNS4 gene, encoding a cytochrome P450 protein, is a novel allele of DWARF11 (D11). A single nucleotide polymorphism (deletion) in the promoter region of GNS4 reduced its expression level in the mutant, leading to reduced grain number and smaller grains. Morphological and cellular analyses suggested that GNS4 positively regulates grain size by promoting cell elongation. Overexpression of GNS4 significantly increased organ size, 1000-grain weight, and panicle size, and subsequently enhanced grain yields in both the Nipponbare and Wuyunjing7 (a high-yielding cultivar) backgrounds. These results suggest that GNS4 is key target gene with possible applications in rice yield breeding.

Conclusion

GNS4 was identified as a positive regulator of grain number and grain size in rice. Increasing the expression level of this gene in a high-yielding rice variety enhanced grain yield. GNS4 can be targeted in breeding programs to increase yields.

     

Carotenoids,Phenolic Profile,Mineral Content and Antioxidant Properties in Flesh and Peel of <Emphasis Type="Italic">Prunus persica</Emphasis> Fruits during Two Maturation Stages

Carotenoids and phenolic profile, antioxidant activity as well as concentrations of selected macronutrients (K, N, Mg, Ca and Na) and micronutrients (Zn, Cu and Mn) in flesh and peel of peach fruit were recorded at two harvest dates. Predominant mineral was potassium, followed by calcium, magnesium and sodium. The concentration of most micronutrients was greater in the peel than in the flesh especially in early season. The concentration of most elements in flesh and peel decreased during fruit maturation. Total carotenoids content varied with respect to the cultivar. β-cryptoxanthin and β-carotene were the major carotenoids in both tissues and flesh contain the lowest amounts. Neochlorogenic acid, chlorogenic acid, catechin, epicatechin, gallic acid, rutin, quercetin-3-O-galactoside, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, were detected in both peel and flesh, with chlorogenic acid and catechin being the predominant components. Peel extracts showed markedly higher antioxidant activities, when estimated by ABTS or DPPH assays, than the flesh counterparts, consistent with the observed higher phenolic content. Overall, total phenolics levels increased at full ripening stage in both peel and flesh. The results found herein provide important data on carotenoids, phenolic and macro- and micronutrient changes during fruit growth, and emphases peach fruit as a potential functional food.     

The Nutritional and Health Benefits of Kiwiberry (<Emphasis Type="Italic">Actinidia arguta</Emphasis>) – a Review

The kiwiberry (Actinidia arguta) is a new product on the market that is enjoying growing consumer acceptance around the world. This widespread interest has created increased demand for identification of the kiwiberry’s nutritional health benefits. Containing over 20 essential nutrients and a range of vitamins, the kiwiberry comes near the top of fruits classed as superfoods. It is one of the richest sources of vitamin C with up to 430 mg/100 g fresh weight (FW) and is considered the richest dietary source of myo-inositol (up to 982 mg/100 g FW). The kiwiberry is also one of the richest sources of lutein (up to 0.93 mg/100 g FW) in commonly consumed fruit. Furthermore, containing up to 1301.1 mg/100 g FW phenolics and significant amounts of the essential minerals of potassium, calcium and zinc, the kiwiberry rates very highly as a ‘healthy food’. The type and number of this fruit’s medicinally promising nutrients have motivated ongoing investigations into its antioxidant, anti-tumour and anti-inflammatory properties. Early research has pointed to the kiwiberry being a very promising treatment for some cancers and health issues involving the gastrointestinal system, hypercholesterolemia and certain cancers. A pharmaceutical composition of A. arguta, A. kolomikta, and A. polygama extracts has already been registered for the prevention and treatment of some immune (inflammatory) mediated diseases, as well as the treatment of some non-allergic inflammatory diseases. This paper reviews and highlights the limited nutritional and therapeutic information currently available on the kiwiberry, a minor fruit possessing such major properties.     

Stir-Frying of Chinese Cabbage and Pakchoi Retains Health-Promoting Glucosinolates

Stir-frying is a cooking method, originating from Asia, in which food is fried in small amount of very hot oil. Nowadays in many other parts of the world stir-frying is a very popular method to prepare vegetables, because it is fast and fried vegetables are tasty. However, the retention of phytochemicals like the health-beneficial glucosinolates in Brassica vegetables is less explored for stir-frying in comparison to other cooking methods. This study investigates the retention of glucosinolates in Chinese cabbage (Brassica rapa ssp. pekinensis) and pakchoi (Brassica rapa ssp. chinensis) as affected by stir-frying at various cooking durations and temperatures. Stir-frying experiments were performed at set pan temperatures ranging from 160 to 250 °C for a duration of 1 to 8 min. Results showed that aliphatic glucobrassicanapin is the most abundant glucosinolate identified in fresh Chinese cabbage and pakchoi, contributing for 48 and 63% of the total glucosinolate content, respectively, followed by glucoiberin and gluconapin. Stir-frying retains the glucosinolates even at the highest temperature applied. Such retention is explained by the quick inactivation of the glucosinolate-hydrolytic enzyme myrosinase during the first minutes of frying, and by the thermal stability of the glucosinolates at those temperature/time conditions. Moreover, due to the absence of a separate water phase, leaching losses did not occur, in contrast to what is observed when boiling Brassica vegetables. These results show that stir-frying may be a suitable health-beneficial cooking option that prevents the loss of glucosinolates.     

Rooting Characteristics of <Emphasis Type="Italic">Solanum chacoense</Emphasis> and <Emphasis Type="Italic">Solanum tuberosum in Vitro</Emphasis>

The precise level of environmental control in vitro may aid in identifying genetically superior plant germplasm for rooting characteristics (RC) linked to increased foraging for plant nitrogen (N). The objectives of this research were to determine the phenotypic variation in root morphological responses of 49 Solanum chacoense (chc), 30 Solanum tuberosum Group Phureja – Solanum tuberosum Group Stenotomum (phu-stn), and three Solanum tuberosum (tbr) genotypes to 1.0 and 0.5 N rate in vitro for 28 d, and identify genotypes with superior RC. The 0.5 N significantly increased density of root length, surface area, and tips. All RC were significantly greater in chc than in either phu-stn or tbr. Based upon clustering on root length, surface area, and volume, the cluster with the greatest rooting values consisted of eight chc genotypes that may be utilized to initiate a breeding program to improve RC in potato.     

Draft Genome Sequence of Potato Dry Rot Pathogen <Emphasis Type="Italic">Fusarium sambucinum</Emphasis> Fckl. F-4

Fusarium sambucinum is one of the most important causal agents that not only cause the dry rot disease of potato tubers in fields and stores worldwide but also capable of producing secondary metabolites toxic for people and animals. Here we present the first draft genome sequence of the strain (F-4) estimated to be around appx. 42.0 Mb. The genome has 12,845 protein coding genes with more than 35,900 exons and gene density of 3.13 per 10Kb. F. sambucinum is evolutionary more close to the F. graminearum among the Fusarium species complex. The genome sequence represents a valuable resource for understanding the pathogenecity and virulence factors, and their evolution within the complex and highly plastic genus Fusarium.     

Marker assisted pyramiding of <Emphasis Type="Italic">Bph6</Emphasis> and <Emphasis Type="Italic">Bph9</Emphasis> into elite restorer line 93–11 and development of functional marker for <Emphasis Type="Italic">Bph9</Emphasis>

Background

The brown planthopper (BPH) has become the most destructive and a serious threat to the rice production in Asia. Breeding the resistant varieties with improved host resistance is the most effective and ecosystem-friendly strategy of BPH biological management. As host resistance was always broken down by the presence of the upgrading BPH biotype, the more resistant varieties with novel resistance genes or pyramiding known identified BPH resistance genes would be needed urgently for higher resistant level and more durability of resistance.

Results

Here, we developed near isogenic lines of Bph9 (NIL-Bph9) by backcrossing elite cultivar 93–11 with Pokkali (harboring Bph9) using marker-assisted selection (MAS). Subsequently, we pyramided Bph6 and Bph9 in 93–11 genetic background through MAS. The resulting Bph6 and Bph9 pyramided line LuoYang69 had stronger antixenotic and antibiosis effects on BPH and exhibited significantly enhanced resistance to BPH than near isogenic lines NIL-Bph6 and NIL-Bph9. LuoYang69 derived hybrids, harboring heterozygous Bph6 and Bph9 genes, also conferred high level of resistance to BPH. Furthermore, LuoYang69 did not affect the elite agronomic traits and rice grain quality of 93–11. The current study also developed functional markers for Bph9. Using functional dominant marker, we screened and evaluated worldwide accessions of rice germplasm. Of the 673 varieties tested, 8 cultivars were identified to harbor functional Bph9 gene.

Conclusion

The development of Bph6 and Bph9 pyramided line LuoYang69 provides valuable resource to develop hybrid rice with highly and durable BPH resistance. The development of functional markers will promote MAS of Bph9. The identified Bph9 containing cultivars can be used as new sources for BPH resistance breeding programs.

     

Root cone angle is enlarged in <Emphasis Type="Italic">docs1</Emphasis> LRR-RLK mutants in rice

Background

The DEFECTIVE IN OUTER CELL LAYER SPECIFICATION 1 (DOCS1) gene belongs to the Leucine-Rich Repeat Receptor-Like Kinase (LRR-RLK) subfamily. It has been discovered few years ago in Oryza sativa (rice) in a screen to isolate mutants with defects in sensitivity to aluminum. The c68 (docs1–1) mutant possessed a nonsense mutation in the C-terminal part of the DOCS1 kinase domain.

Findings

We have generated a new loss-of-function mutation in the DOCS1 gene (docs1–2) using the CRISPR-Cas9 technology. This new loss-of-function mutant and docs1–1 present similar phenotypes suggesting the original docs1–1 was a null allele. Besides the aluminum sensitivity phenotype, both docs1 mutants shared also several root phenotypes described previously: less root hairs and mixed identities of the outer cell layers. Moreover, our new results suggest that DOCS1 could also play a role in root cap development. We hypothesized these docs1 root phenotypes may affect gravity responses. As expected, in seedlings, the early gravitropic response was delayed. Furthermore, at adult stage, the root gravitropic set angle of docs1 mutants was also affected since docs1 mutant plants displayed larger root cone angles.

Conclusions

All these observations add new insights into the DOCS1 gene function in gravitropic responses at several stages of plant development.

     

Association of Potato Psyllid (<Emphasis Type="Italic">Bactericera cockerelli</Emphasis>; Hemiptera: Triozidae) with <Emphasis Type="Italic">Lycium</Emphasis> spp. (Solanaceae) in Potato Growing Regions of Washington,Idaho, and Oregon

Potato psyllid, Bactericera cockerelli (?ulc), causes economic damage to potato crops throughout the major potato growing regions of western North America. When cultivated crops are not available, potato psyllid often occurs on non-crop hosts. In the southern U.S. and northern Mexico, native species of Lycium (Solanaceae) are important non-crop hosts for the psyllid. We determined whether Old World species of Lycium now widespread in the Pacific Northwest are reservoirs of potato psyllid in this growing region. We examined Lycium spp. across a wide geographic region in Washington, Oregon, and Idaho at irregular intervals during three growing seasons. Potato psyllids were present at all locations. To determine whether Lycium is also a host during intervals of the year in which the potato crop is not available, we monitored a subset of these sites over the entire year. Six sites were monitored at 1- to 3-week intervals from June 2014 to June 2016. Psyllids were present on Lycium throughout the year at all sites, including during winter, indicating that Lycium is also a host when the potato crop is seasonally not available. Psyllid populations included a mixture of Northwestern and Western haplotypes. We observed well-defined spring and fall peaks in adult numbers, with peaks separated by long intervals in which psyllid numbers were very low. Seasonal patterns in psyllid numbers on these non-native Lycium hosts were very similar to what has been observed on native Lycium in the desert southwest region of the U.S. Our findings demonstrate that potato psyllid associates with Lycium across a broad geographic region within the Pacific Northwest. These results will assist in predicting sources of potato psyllid colonizing potatoes in this important growing region.     

Dissecting combining ability effect in a rice NCII-III population provides insights into heterosis in <Emphasis Type="Italic">indica-japonica</Emphasis> cross

Background

Combining ability is a measure for selecting elite parents that make the highest contributions to hybrid performance. However, the genetic bases of combining ability and how they contributed to heterosis is seldomly known.

Results

We constructed a both NCII and NCIII population derived from an indica-japonica cross to study the relationship among parental performance, combining ability and hybrid performance of 11 agronomic traits. Among them, specific combining ability is more important to grain yield than parental performance and general combining ability. We performed linkage analyses to phenotypic values and combining ability of all 11 traits in Doubled haploid lines and its two backcross populations and identified 108 QTLs in total. Among these QTLs, four known loci, Sd1, Ghd7, Ghd8 and DEP1 contribute a lot to GCA effects of agronomic traits except grain yield and seed setting rate. Three QTLs, Ghd8, S5 and qS12, contribute a lot to SCA effects of grain yield and present overdominace.

Conclusions

Our study provides insights into the genetic bases of combining ability and heterosis and will promote the improvements of indica-japonica hybrid breeding.

     

Identification of resistant germplasm containing novel resistance genes at or tightly linked to the <Emphasis Type="Italic">Pi2/9</Emphasis> locus conferring broad-spectrum resistance against rice blast

Background

The rice Pi2/9 locus harbors multiple resistance (R) genes each controlling broad-spectrum resistance against diverse isolates of Magnaporthe oryzae, a fungal pathogen causing devastating blast disease to rice. Identification of more resistance germplasm containing novel R genes at or tightly linked to the Pi2/9 locus would promote breeding of resistance rice cultivars.

Results

In this study, we aim to identify resistant germplasm containing novel R genes at or tightly linked to the Pi2/9 locus using a molecular marker, designated as Pi2/9-RH (Pi2/9 resistant haplotype), developed from the 5′ portion of the Pi2 sequence which was conserved only in the rice lines containing functional Pi2/9 alleles. DNA analysis using Pi2/9-RH identified 24 positive lines in 55 shortlisted landraces which showed resistance to 4 rice blast isolates. Analysis of partial sequences of the full-length cDNAs of Pi2/9 homologues resulted in the clustering of these 24 lines into 5 haplotypes each containing different Pi2/9 homologues which were designated as Pi2/9-A5, ?A15, ?A42, ?A53, and -A54. Interestingly, Pi2/9-A5 and Pi2/9-A54 are identical to Piz-t and Pi2, respectively. To validate the association of other three novel Pi2/9 homologues with the blast resistance, monogenic lines at BC₃F₃ generation were generated by marker assisted backcrossing (MABC). Resistance assessment of the derived monogenic lines in both the greenhouse and the field hotspot indicated that they all controlled broad-spectrum resistance against rice blast. Moreover, genetic analysis revealed that the blast resistance of these three monogenic lines was co-segregated with Pi2/9-RH, suggesting that the Pi2/9 locus or tightly linked loci could be responsible for the resistance.

Conclusion

The newly developed marker Pi2/9-RH could be used as a potentially diagnostic marker for the quick identification of resistant donors containing functional Pi2/9 alleles or unknown linked R genes. The three new monogenic lines containing the Pi2/9 introgression segment could be used as valuable materials for disease assessment and resistance donors in breeding program.

     

<Emphasis Type="Italic">OsLAP6/OsPKS1</Emphasis>, an orthologue of <Emphasis Type="Italic">Arabidopsis PKSA/LAP6</Emphasis>, is critical for proper pollen exine formation

Background

Male fertility is crucial for rice yield, and the improvement of rice yield requires hybrid production that depends on male sterile lines. Although recent studies have revealed several important genes in male reproductive development, our understanding of the mechanisms of rice pollen development remains unclear.

Results

We identified a rice mutant oslap6 with complete male sterile phenotype caused by defects in pollen exine formation. By using the MutMap method, we found that a single nucleotide polymorphism (SNP) variation located in the second exon of OsLAP6/OsPKS1 was responsible for the mutant phenotype. OsLAP6/OsPKS1 is an orthologous gene of Arabidopsis PKSA/LAP6, which functions in sporopollenin metabolism. Several other loss-of-function mutants of OsLAP6/OsPKS1 generated by the CRISPR/Cas9 genomic editing tool also exhibited the same phenotype of male sterility. Our cellular analysis suggested that OsLAP6/OsPKS1 might regulate pollen exine formation by affecting bacula elongation. Expression examination indicated that OsLAP6/OsPKS1 is specifically expressed in tapetum, and its product is localized to the endoplasmic reticulum (ER). Protein sequence analysis indicated that OsLAP6/OsPKS1 is conserved in land plants.

Conclusions

OsLAP6/OsPKS1 is a critical molecular switch for rice male fertility by participating in a conserved sporopollenin precursor biosynthetic pathway in land plants. Manipulation of OsLAP6/OsPKS1 has potential for application in hybrid rice breeding.