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Abstract The transformation and gene editing of the woody species kiwifruit are difficult and time-consuming. The fast and marker-free genetic modification system for kiwifruit has not been developed yet. Here, we establish a rapid and efficient marker-free transformation and gene editing system mediated by Agrobacterium rhizogenes for kiwifruit. Moreover, a removing-root-tip method was developed to significantly increase the regeneration efficiency of transgenic hairy roots. Through A. rhizogenes-mediated CRISPR/Cas9 gene editing, the editing efficiencies of CEN4 and AeCBL3 achieved 55 and 50%, respectively. And several homozygous knockout lines for both genes were obtained. Our method has been successfully applied in the transformation of two different species of kiwifruit (Actinidia chinensis ‘Hongyang’ and A.eriantha ‘White’). Next, we used the method to study the formation of calcium oxalate (CaOx) crystals in kiwifruit. To date, little is known about how CaOx crystal is formed in plants. Our results indicated that AeCBL3 overexpression enhanced CaOx crystal formation, but its knockout via CRISPR/Cas9 significantly impaired crystal formation in kiwifruit. Together, we developed a fast maker-free transformation and highly efficient CRISPR-Cas9 gene editing system for kiwifruit. Moreover, our work revealed a novel gene mediating CaOx crystal formation and provided a clue to elaborate the underlying mechanisms. Graphical abstract

We previously built a sample of 14,012 extremely variable quasars (EVQs) based on Sloan Digital Sky Survey (SDSS) and Pan-STARRS1 photometric observations. In this work we present the spectral fitting to their SDSS spectra and study the spectral variation in 1259 EVQs with multiepoch SDSS spectra (after prudently excluding spectra with potentially unreliable spectroscopic photometry). We find a clear “bluer-when-brighter” trend in EVQs, consistent with previous findings of normal quasars and active galactic nuclei. We detect significant intrinsic Baldwin effect (iBeff, i.e., smaller line equivalent width at higher continuum flux in individual active galactic nuclei) in the broad Mg ii and C iv lines of EVQs. Meanwhile, no systematical iBeff is found for the broad H β line, which could be attributed to strong host contamination at longer wavelengths. Remarkably, by comparing the iBeff slope of EVQs with archived changing-look quasars, we show that the changing-look quasars identified in the literature are most likely a biased (due to its definition) subpopulation of EVQs, rather than a distinct population of quasars. We also found no significant broad line breathing of H β , Mg ii , or C iv , suggesting the broad line breathing in quasars may disappear at longer timescales (∼3000 days).

Summary: The chloroplast is the main organelle for stress-induced production of reactive oxygen species (ROS). However, how chloroplastic ROS homeostasis is maintained under salt stress is largely unknown. We show that EGY3, a gene encoding a chloroplast-localized protein, is induced by salt and oxidative stresses. The loss of EGY3 function causes stress hypersensitivity while EGY3 overexpression increases the tolerance to both salt and chloroplastic oxidative stresses. EGY3 interacts with chloroplastic Cu/Zn-SOD2 (CSD2) and promotes CSD2 stability under stress conditions. In egy3-1 mutant plants, the stress-induced CSD2 degradation limits H2O2 production in chloroplasts and impairs H2O2-mediated retrograde signaling, as indicated by the decreased expression of retrograde-signal-responsive genes required for stress tolerance. Both exogenous application of H2O2 (or APX inhibitor) and CSD2 overexpression can rescue the salt-stress hypersensitivity of egy3-1 mutants. Our findings reveal that EGY3 enhances the tolerance to salt stress by promoting the CSD2 stability and H2O2-mediated chloroplastic retrograde signaling.