Estimation of leaf nutrient concentration from hyperspectral reflectance in Eucalyptus using partial least squares regression

Leaf hyperspectral reflectance has been used to estimate nutrient concentrations in plants in narrow bands of the electromagnetic spectrum. The aim of this study was to estimate leaf nutrient concentrations using leaf hyperspectral reflectance and verify the variable selection methods using the partial least squares regression (PLSR). Two studies were carried out using stands with Eucalyptus clones. Study I was established in Eucalyptus stands with three clones, classifying leaves into five colour patterns using the Munsell chart for plant tissues. Immediately after leaf collection, leaf reflectance was read and the chemical analysis was performed. Study II was carried out in commercial clonal stands of Eucalyptus performing the same leaf sampling and chemical analysis as used in Study I. All leaf reflectance spectra were smoothed and three more pre-processing procedures were applied. In addition, three methods of PLSR were tested. The first derivative was more accurate for predicting nitrogen ( Rcv2=0.95 ), phosphorous ( Rcv2=0.93 ), and sulphur concentration ( Rcv2=0.85 ). The estimates for concentrations of calcium ( Rcv2=0.81 ), magnesium ( Rcv2=0.22 ), and potassium ( Rcv2=0.76 ) were more accurate using the logarithm transformation. Only the estimates for iron concentrations were performed with higher accuracy ( Rcv2=0.35 ) using the smoothed reflectance. The copper concentrations were more accurate ( Rcv2=0.78 ) using the logarithm transformation. Concentrations of boron ( Rcv2=0.68 ) and manganese ( Rcv2=0.79 ) were more accurate using the first derivative, while zinc ( Rcv2=0.31 ) concentration was most accurate using the second derivative.

Estimation of leaf nutrient concentration from hyperspectral reflectance in Eucalyptus using partial least squares regression

Leaf hyperspectral reflectance has been used to estimate nutrient concentrations in plants in narrow bands of the electromagnetic spectrum. The aim of this study was to estimate leaf nutrient concentrations using leaf hyperspectral reflectance and verify the variable selection methods using the partial least squares regression (PLSR). Two studies were carried out using stands with Eucalyptus clones. Study I was established in Eucalyptus stands with three clones, classifying leaves into five colour patterns using the Munsell chart for plant tissues. Immediately after leaf collection, leaf reflectance was read and the chemical analysis was performed. Study II was carried out in commercial clonal stands of Eucalyptus performing the same leaf sampling and chemical analysis as used in Study I. All leaf reflectance spectra were smoothed and three more pre-processing procedures were applied. In addition, three methods of PLSR were tested. The first derivative was more accurate for predicting nitrogen ( Rcv2=0.95 ), phosphorous ( Rcv2=0.93 ), and sulphur concentration ( Rcv2=0.85 ). The estimates for concentrations of calcium ( Rcv2=0.81 ), magnesium ( Rcv2=0.22 ), and potassium ( Rcv2=0.76 ) were more accurate using the logarithm transformation. Only the estimates for iron concentrations were performed with higher accuracy ( Rcv2=0.35 ) using the smoothed reflectance. The copper concentrations were more accurate ( Rcv2=0.78 ) using the logarithm transformation. Concentrations of boron ( Rcv2=0.68 ) and manganese ( Rcv2=0.79 ) were more accurate using the first derivative, while zinc ( Rcv2=0.31 ) concentration was most accurate using the second derivative.(AU)

Identification of the CAD gene from Eucalyptus urophylla GLU4 and its functional analysis in transgenic tobacco.

Cinnamyl alcohol dehydrogenase (CAD) catalyzes the final step in lignin biosynthesis. The genus Eucalyptus belongs to the family Myrtaceae, which is the main cultivated species in China. Eucalyptus urophylla GLU4 (GLU4) is widely grown in Guangxi. It is preferred for pulping because of its excellent cellulose content and fiber length. Based on GLU4 and CAD gene expression, a Eucalyptus variety low in lignin content should be obtained using transgenic technology, which could reduce the cost of pulp and improve the pulping rate, and have favorable prospects for application. However, the role and function of CAD in GLU4 is still unclear. In the present study, EuCAD was cloned from GLU4 and identified using bioinformatic tools. Subsequently, in order to evaluate its impact on lignin synthesis, a full-length EuCAD RNAi vector was constructed, and transgenic tobacco was obtained via Agrobacterium-mediated transformation. A significant decrease in CAD expression and lignin content in transgenic tobacco demonstrated a key role for EuCAD in lignin biosynthesis and established a regulatory role for RNAi. In our study, the direct molecular basis of EuCAD expression was determined, and the potential regulatory effects of this RNAi vector on lignin biosynthesis in E. urophylla GLU4 were demonstrated. Our results provide a theoretical basis for the study of lignin biosynthesis in Eucalyptus.

Rescue of syringyl lignin and sinapate ester biosynthesis in Arabidopsis thaliana by a coniferaldehyde 5-hydroxylase from Eucalyptus globulus.

KEY MESSAGE: The gene coding for F5H from Eucalyptus globulus was cloned and used to transform an f5h -mutant of Arabidopsis thaliana , which was complemented, thus verifying the identity of the cloned gene. Coniferaldehyde 5-hydroxylase (F5H; EC 1.14.13) is a cytochrome P450-dependent monooxygenase that catalyzes the 5-hydroxylation step required for the production of syringyl units in lignin biosynthesis. The Eucalyptus globulus enzyme was characterized in vitro, and results showed that the preferred substrates were coniferaldehyde and coniferyl alcohol. Complementation experiments demonstrated that both cDNA and genomic constructs derived from F5H from E. globulus under the control of the cinnamate 4-hydroxylase promoter from Arabidopsis thaliana, or a partial F5H promoter from E. globulus, can rescue the inability of the A. thaliana fah1-2 mutant to accumulate sinapate esters and syringyl lignin. E. globulus is a species widely used to obtain products that require lignin removal, and the results suggest that EglF5H is a good candidate for engineering efforts aimed at increasing the lignin syringyl unit content, either for kraft pulping or biofuel production.

Drought and salt tolerance enhancement of transgenic Arabidopsis by overexpression of the vacuolar pyrophosphatase 1 (EVP1) gene from Eucalyptus globulus.

Vacuolar solute accumulation has been shown to be a mechanism by which plants are capable of increasing drought and salt tolerance. The exposure of plants to NaCl induces H+ transport into the vacuole by specialized pumps. One of them corresponds to the vacuolar H+-pyrophosphatase, which generates a H+ gradient across the vacuolar membrane. In our laboratory we isolated the first cDNA sequence of a vacuolar pyrophosphatase type I (EVP1) from Eucalyptus globulus. Using real-time PCR we confirmed that EVP1 participates in Eucalyptus plants' response to drought and salt stress through an ABA independent pathway. Additionally, the overexpression of EVP1 in transgenic Arabidopsis resulted in an enhancement of drought and salt tolerance. Interestingly we established that the transgenic plants had a higher number of root hairs, which may have a positive effect on the plant's response to drought and salt stress. These results suggest that EVP1 plays an active role in abiotic stress tolerance in E. globulus, and that it may be potentially used to enhance drought and stress tolerance of plants.