Identification of MaWRKY40 and MaDLO1 as Effective Marker Genes for Tracking the Salicylic Acid-Mediated Immune Response in Bananas.

Bananas are among the world's most important cash and staple crops but are threatened by various devastating pathogens. The phytohormone salicylic acid (SA) plays a key role in the regulation of plant immune response. Tracking the expression of SA-responsive marker genes under pathogen infection is important in pathogenesis elucidation. However, the common SA-responsive marker genes are not consistently induced in different banana cultivars or different organs. Here, we conducted transcriptome analysis for SA response of a banana cultivar, 'Pei-Chiao' (Cavendish, AAA genome), and identified three genes, MaWRKY40, MaWRKY70, and Downy Mildew Resistant 6 (DMR6)-Like Oxygenase 1 (MaDLO1) that are robustly induced upon SA treatment in both the leaves and roots. Consistent induction of these three genes by SA treatment was also detected in both the leaves and roots of bananas belonging to different genome types such as 'Tai-Chiao No. 7' (Cavendish, AAA genome), 'Pisang Awak' (ABB genome), and 'Lady Finger' (AA genome). Furthermore, the biotrophic pathogen cucumber mosaic virus elicited the expression of MaWRKY40 and MaDLO1 in infected leaves of susceptible cultivars. The hemibiotrophic fungal pathogen Fusarium oxysporum f. sp. cubense tropical race 4 (TR4) also consistently induced the expression of MaWRKY40 and MaDLO1 in the infected roots of the F. oxysporum f. sp. cubense TR4-resistant cultivar. These results indicate that MaWRKY40 and MaDLO1 can be used as reliable SA-responsive marker genes for the study of plant immunity in banana. Revealing SA-responsive marker genes provides a stepping stone for further studies in banana resistance to pathogens.

Iron is involved in the maintenance of circadian period length in Arabidopsis.

The homeostasis of iron (Fe) in plants is strictly regulated to maintain an optimal level for plant growth and development but not cause oxidative stress. About 30% of arable land is considered Fe deficient because of calcareous soil that renders Fe unavailable to plants. Under Fe-deficient conditions, Arabidopsis (Arabidopsis thaliana) shows retarded growth, disordered chloroplast development, and delayed flowering time. In this study, we explored the possible connection between Fe availability and the circadian clock in growth and development. Circadian period length in Arabidopsis was longer under Fe-deficient conditions, but the lengthened period was not regulated by the canonical Fe-deficiency signaling pathway involving nitric oxide. However, plants with impaired chloroplast function showed long circadian periods. Fe deficiency and impaired chloroplast function combined did not show additive effects on the circadian period, which suggests that plastid-to-nucleus retrograde signaling is involved in the lengthening of circadian period under Fe deficiency. Expression pattern analyses of the central oscillator genes in mutants defective in CIRCADIAN CLOCK ASSOCIATED1/LATE ELONGATED HYPOCOTYL or GIGANTEA demonstrated their requirement for Fe deficiency-induced long circadian period. In conclusion, Fe is involved in maintaining the period length of circadian rhythm, possibly by acting on specific central oscillators through a retrograde signaling pathway.

Arabidopsis SUMO E3 ligase SIZ1 is involved in excess copper tolerance.

The reversible conjugation of the small ubiquitin-like modifier (SUMO) to protein substrates occurs as a posttranslational regulatory process in eukaryotic organisms. In Arabidopsis (Arabidopsis thaliana), several stress-responsive SUMO conjugations are mediated mainly by the SUMO E3 ligase SIZ1. In this study, we observed a phenotype of hypersensitivity to excess copper in the siz1-2 and siz1-3 mutants. Excess copper can stimulate the accumulation of SUMO1 conjugates in wild-type plants but not in the siz1 mutant. Copper accumulated to a higher level in the aerial parts of soil-grown plants in the siz1 mutant than in the wild type. A dramatic difference in copper distribution was also observed between siz1 and wild-type Arabidopsis treated with excess copper. As a result, the shoot-to-root ratio of copper concentration in siz1 is nearly twice as high as that in the wild type. We have found that copper-induced Sumoylation is involved in the gene regulation of metal transporters YELLOW STRIPE-LIKE 1 (YSL1) and YSL3, as the siz1 mutant is unable to down-regulate the expression of YSL1 and YSL3 under excess copper stress. The hypersensitivity to excess copper and anomalous distribution of copper observed in the siz1 mutant are greatly diminished in the siz1ysl3 double mutant and slightly in the siz1ysl1 double mutant. These data suggest that SIZ1-mediated sumoylation is involved specifically in copper homeostasis and tolerance in planta.

Photosynthetic and Morphological Responses of Sacha Inchi (Plukenetia volubilis L.) to Waterlogging Stress.

Sacha inchi (Plukenetia volubilis L.) is an important oilseed crop that is rich in fatty acids and protein. Climate-change-related stresses, such as chilling, high temperature, and waterlogging can cause severe production loss in this crop. In this study, we investigated the photosynthetic responses of sacha inchi seedlings to short-term waterlogging and their morphological changes after long-term waterlogging stress. Sacha inchi CO2 uptake, stomatal conductance, and transpiration rate are affected by temperature and light intensity. The seedlings had a high CO2 uptake (>10 µmol m-2s-1) during the daytime (08:00 to 15:00), and at 32 and 36 °C. At 32 °C, CO2 uptake peaked at irradiations of 1000 and 1500 µmol m-2s-1, and plants could still perform photosynthesis at high-intensity radiation of 2000-3000 µmol m-2s-1. However, after 5 days of waterlogging (5 DAF) sacha inchi seedlings significantly reduced their photosynthetic ability. The CO2 uptake, stomatal conductance, Fv/Fm, ETR, and qP, etc., of the susceptible genotypes, were significantly decreased and their wilting percentage was higher than 50% at 5 DAF. This led to a higher wilting percentage at 7 days post-recovery. Among the four lines assessed, Line 27 had a high photosynthetic capability and showed the best waterlogging tolerance. We screened many seedlings for long-term waterlogging tolerance and discovered that some seedlings can produce adventitious roots (AR) and survive after two weeks of waterlogging. Hence, AR could be a critical morphological adaptation to waterlogging in this crop. In summary, these results suggest that improvement in waterlogging tolerance has considerable potential for increasing the sustainable production of sacha inchi.

Arabidopsis IRT3 is a zinc-regulated and plasma membrane localized zinc/iron transporter.

ZIP transporters (ZRT, IRT-like proteins) are involved in the transport of iron (Fe), zinc (Zn) and other divalent metal cations. The expression of IRT3, a ZIP transporter, is higher in the Zn/cadmium (Cd) hyperaccumulator Arabidopsis halleri than is that of its ortholog in Arabidopsis thaliana, which implies a positive association of its expression with Zn accumulation in A. halleri. IRT3 genes from both A. halleri and A. thaliana functionally complemented the Zn uptake mutant Spzrt1 in Schizosaccharomyces pombe; and Zn uptake double mutant zrt1zrt2, Fe-uptake mutant fet3fet4 and conferred Zn and Fe uptake activity in Saccharomyces cerevisiae. By contrast, the manganese (Mn) uptake mutant smf1 phenotypes were not rescued. Insufficient Cd uptake for toxicity was found. Expression of IRT3-green fluorescent protein (GFP) fusion proteins in Arabidopsis root protoplasts indicated localization of both IRT3 proteins in the plasma membrane. Overexpressing AtIRT3 in A. thaliana led to increased accumulation of Zn in the shoot and Fe in the root of transgenic lines. Therefore, IRT3 functions as a Zn and Fe-uptake transporter in Arabidopsis.

Patterned growth of ZnO nanostructures based on the templation of plant cell walls.

A new and general approach based on vapor-phase transport technique using Au-coated plant cell walls has been developed to synthesize patterned ZnO nanostructures. Nanowires, nanodendrites and nanotowers were fabricated by adsorption of different metallic ions on plant cell walls. It is shown that plant cell wall can serve as a well-defined template to grow patterned nanostructures. Using transmission electron microscope and Raman spectroscopy, the structural characteristic of the nanostructures were investigated, exhibiting good crystallinity and hexagonal symmetry of the nanomaterials. Quite interestingly, the shape of the nanostructures can be controlled by the metallic ions adsorbed on plant cell walls. Without metallic ions, a homogeneous distribution of nanowires was obtained. On the other hand, with Ni+2 or Fe+3 ions, nanodendrites and nanotowers were observed, respectively. Our approach provides a low cost method that opens up new possibilities for the growth of patterned nanomaterials with desired shapes.

Alternative functions of Arabidopsis Yellow Stripe-Like3: from metal translocation to pathogen defense.

Yellow stripe-like1 (YSL1) and YSL3 are involved in iron (Fe) and copper (Cu) translocation. Previously, we reported that upregulation of YSL1 and YSL3 under excess Cu caused high accumulation of Cu in the siz1 mutant, impaired in small ubiquitin-like modifier (SUMO) E3 ligase. Interestingly, the siz1 mutant contains high levels of salicylic acid (SA), involved in plant defense against biotrophic pathogens. In this study, we found that YSL1 and YSL3 were upregulated by SA. SA-regulated YSL3 but not YSL1 depended on nonexpressor of PR1 (NPR1). Susceptibility to the pathogen Pseudomonas syringe pv. tomato (Pst) DC3000 was greater for ysl3 than the wild type. Also, during Pst DC3000 infection, YSL3 was positively regulated by SA signaling through NPR1 and the upregulation was enhanced in the coi1 mutant that defective in the jasmonic acid (JA) receptor, coronatine insensitive1. This line of evidence indicates that the regulation of YSL3 is downstream of SA signaling and interplays with JA signaling for involvement in pathogen-induced defense. We provide new insights into the biological function of the metal transporter YSL3 in plant pathogen defense.

Effect of Cu content on the activity of Cu/ZnSOD1 in the Arabidopsis SUMO E3 ligase siz1 mutant.

In a previous study, we found copper (Cu) accumulated to a higher level in the aerial parts of soil-grown plants of the SUMO E3 ligase siz1 mutant than in those of the wild type. Here, we found that all superoxide dismutase (SOD) isoforms, such as FeSOD, MnSOD and different types of Cu/ZnSOD, were more active in the siz1 mutant than in the wild type under normal growth conditions. We further examined the expression and enzymatic activity of Cu/ZnSOD1 (CSD1) in shoots of the siz1 mutant under excess Cu. Shoot CSD1 protein level and activity were reduced in siz1 with excess Cu but induced in the wild type. SIZ1-dependent SUMOylation may be involved in maintaining CSD1 protein stability or repelling a feedback regulation under Cu stress.

A new method to measure leaf age: Leaf measuring-interval index.

We propose a new method, the leaf measuring-interval index (LMI), to estimate leaf age in morphological and physiological studies of leaves. When the plastochron, the interval between the initiation of successive leaves, is constant, the well-known leaf plastochron index (LPI) provides a robust measure of leaf age. When the duration of the plastochron is not uniform, however, we show that the LPI can (in simulations) and does (with actual data) turn variation in duration of the plastochron into variance about the regression estimates of leaf growth curves. The method we present in this paper, the LMI, is plastochron independent. This new method is particularly suited, therefore, for studies of plants growing in natural environments rather than in controlled growth facilities where the assumptions of the LPI method can be met.