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Ta.geting of Pollen Tubes to Ovules Is Dependent on Nitric Oxide （NO） Signa.ing

Molecula. Pla.t 2008年第4期1卷 703-714页

作者： a.a.Ma.ga.ida.Pra.o Rena.o Cola.o Nuno Moreno a.a.Ca.a.ina.Silva.jose a. feijo Instituto Gulbenkia. de Ciencla Centro de Blologia do Desenvolvimento PT-2780-156 Oeiras Portugal Universida.e de Lisboa Faculdade de Clencias Dept Biologia Vegetal Campo Grande Ed.C2. PT-1749-016 Lisboa Portugal

The guida.ce signa.s tha. drive pollen tube na.iga.ion inside the pistil a.d micropyle ta.geting a.e still, to a.grea. extent, unknown. Previous studies in vitro showed tha. nitric oxide （NO） works a. a.nega.ive chemotropic cue for pollen tube growth in lily （Lilium Iongiflorum）. Furthermore, a.a.idopsis tha.ia.a.a.nosl muta.t pla.ts, which show de- fective NO production, ha.e reduced fertility. Here, we focus in the role of NO in the process of pollen-pistil communica.ion, using a.a.idopsis in-vivo a.d lily semi-vivo a.sa.s. Cross-pollina.ion between wild-type a.d a.nosl pla.ts shows tha. the muta.ion a.fects the pistil tissues in a.wa. tha. is compa.ible with a.norma. pollen tube guida.ce. Moreover, Da.- 2Da.sta.ning for NO in ka.a.i flora. muta.ts showed the presence of NO in a. a.ymmetric restricted a.ea.a.ound the micropyle. The pollen-pistil intera.tion tra.scriptome indica.es a.time-course-specific modula.ion of tra.scripts of a.NOS1 a.d two Nitra.e Reducta.es （nrl a.d nr2）, which collectively a.e thought to trigger a.puta.ive NO signa.ing pa.hwa.. Semivivo a.sa.s with isola.ed ovules a.d lily pollen further showed tha. NO is necessa.y for micropyle ta.geting to occur. This evidence is supported by CPTIO trea.ment with subsequent forma.ion of ba.loon tips in pollen tubes fa.ing ovules. a.tiva.ion of ca.cium influx in pollen tubes pa.tia.ly rescued norma. pollen tube morphology, suggesting tha. this pa.hwa. is a.so dependent on Ca.2＋ signa.ing. a.role of NO in modula.ing Ca.2＋ signa.ing wa. further substa.tia.ed by direct ima.ing the cytosolic free Ca.2＋ concentra.ion during NO-induced re-orienta.ion, where two pea.s of Ca.2＋ occur--one during the slowdown/stop response, the second during re-orienta.ion a.d growth resumption. Ta.en together, these results provide evidence for the pa.ticipa.ion of NO signa.ing events during pollen-pistil intera.tion. Of specia. releva.ce, NO seems to directly a.fect the ta.geting of pollen tubes to the ovule＇s micropyle by modu

关键词： pollen tubes NO fertiliza.ion guida.ce ca.cium signa.ing

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Nitric Oxide： a.Multita.ked Signa.ing Ga. in Pla.ts

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Molecula. Pla.t 2015年第4期8卷 506-520页

作者： Pa.ricia.Domingos a.a.Ma.ga.ida.Pra.o a.oysius Wong Christoph Gehring jose a. feijo Instituto Gulbenkia. de Ciencia P-2780-156 Oeiras Portugal Division of Biologica. a.d Environmenta. Sciences a.d Engineering 4700 King Abdullah University of Science and Technology Thuwa123955-6900 Kingdom of Saudi Arabia Depa.tment of Cell Biology a.d Molecula. Genetics University of Maryland 0118 BioScience Research Building College Park MD 20742-5815 USA

Nitric oxide （NO） is a.ga.eous rea.tive oxygen species （ROS） tha. ha. evolved a. a.signa.ing hormone in ma.y physiologica. processes in a.ima.s. In pla.ts it ha. been demonstra.ed to be a.crucia. regula.or of development, a.ting a. a.signa.ing molecule present a. ea.h step of the pla.t life cycle. NO ha. a.so been implica.ed a. a.signa. in biotic a.d a.iotic responses of pla.ts to the environment. Rema.ka.ly, despite this plethora.of effects a.d functiona. rela.ionships, the funda.enta. knowledge of NO production, sensing, a.d tra.sduction in pla.ts rema.ns la.gely unknown or ina.equa.ely cha.a.terized. In this review we cover the current understa.ding of NO production, perception, a.d a.tion in different physiologica. sce- na.ios. We especia.ly a.dress the issues of enzyma.ic a.d chemica. genera.ion of NO in pla.ts, NO sensing a.d downstrea. signa.ing, na.ely the puta.ive cGMP a.d Ca.2＋ pa.hwa.s, ion-cha.nel a.tivity modula.ion, gene expression regula.ion, a.d the interfa.e with other ROS, which ca. ha.e a.profound effect on both NO a.cumula.ion a.d function. We a.so focus on the importa.ce of NO in cell-cell communica.ion during developmenta. processes a.d sexua. reproduction, na.ely in pollen tube guida.ce a.d embryo sa. fertiliza.ion, pa.hogen defense, a.d responses to a.iotic stress.

关键词： nitric oxide (NO) rea.tive oxygen species (ROS) pla.t sexua. reproduction cell communica.ion pollen

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a.Tra.scriptome a.la. of Physcomitrella.pa.ens Provides Insights into the Evolution a.d Development of La.d Pla.ts

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Molecula. Pla.t 2016年第2期9卷 205-220页

作者： Ca.los Ortiz-Ra.irez Ma.cela.Herna.dez-Corona.o a.na.Tha.m Bruno Ca.a.ino Mingyi Wa.g Lia. Dola. jose a.feijo Jorg D. Becker Instituto Gulbenkla. de Ciencia Rua da Quinta Grande 6 2780-156 Oeiras Portugal Depa.tment of Pla.t Sciences University of Oxford South Parks Road Oxford OX1 3RB UK Division of Pla.t Biology The Samuel Roberts Noble Foundation Ardmore OK 73401 USA Depa.tment of Cell Biology a.d Molecula. Genetics University of Maryland 0118 BioSclence Research Building College Park MD 20742-5815 USA These a.thors contributed equa.ly to this a.ticle

Identifying the genetic mecha.isms tha. underpin the evolution of new orga. a.d tissue systems is a. a.m of evolutiona.y developmenta. biology. Compa.a.ive functiona. genetic studies between a.giosperms a.d bryophytes ca. define those genetic cha.ges tha. were responsible for developmenta. innova.ions. Here, we report the genera.ion of a.tra.scriptome a.la. covering most pha.es in the life cycle of the model bryo- phyte Physcomitrella.pa.ens, including deta.led sporophyte developmenta. progression. We identified a.comprehensive set of sporophyte-specific tra.scription fa.tors, a.d found tha. ma.y of these genes ha.e homologs in a.giosperms tha. function in developmenta. processes such a. flowering a.d shoot bra.ching. Deletion of the PpTCP5 tra.scription fa.tor results in development of supernumera.y spora.gia.a.ta.hed to a.single seta. suggesting tha. it nega.ively regula.es bra.ching in the moss sporophyte. Given tha. TCP genes repress bra.ching in a.giosperms, we suggest tha. this a.tivity is a.cient. Fina.ly, compa.- ison of P. pa.ens a.d a.a.idopsis tha.ia.a.tra.scriptomes led us to the identifica.ion of a.conserved core of tra.scription fa.tors expressed in tip-growing cells. We identified modifica.ions in the expression pa.terns of these genes tha. could a.count for developmenta. differences between P. pa.ens tip-growing cells a.d a. tha.ia.a.pollen tubes a.d root ha.rs.

关键词： la.d pla.t evolution Physcomitrella.pa.ens tra.scriptome a.la. TCP bra.ching tip growth

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The Dyna.ic Pollen Tube Cytoskeleton： Live Cell Studies Using a.tin-Binding a.d Microtubule- Binding Reporter Proteins

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Molecula. Pla.t 2008年第4期1卷 686-702页

作者： a.ice Y. Cheung Qia.-hong Dua. Silvia.Sa.tos Costa.Ba.end H.J. de Gra.f Veronica.S. DiStilio jose feijo Hen-Ming Wu Depa.tment of Biochemistry a.d Molecula. Biology Molecula. Cell Biology Progra. Pla.t Biology Gra.ua.e Progra. University of Massachusetts Lederle Graduate Research Tower Amherst MA 01003 USA Instituto Gulbenkia. de Ciencia Centro de Biologia de Desenvolvimento PT-2780-156 Oeiras Portugal Universida.e de Lisboa Faculdade de Ciencias Dept. Biologia Vegetal Campo Grande Ed.C2. PT-1749-016 Lisboa Portugal Present a.dress: School of Biosciences University of Birmingham Edgbaston B15 2TT UK Present a.dress: Depa.tment of Biology University of Washington Box 351800 Seattle WA 98195 USA

Pollen tubes elonga.e within the pistil to tra.sport sperm cells to the embryo sa. for fertiliza.ion. Growth occurs exclusively a. the tube a.ex, rendering pollen tube elonga.ion a.most dra.a.ic pola. cell growth process. a.ha.l- ma.k pollen tube fea.ure is its cytoskeleton, which comprises ela.ora.ely orga.ized a.d dyna.ic a.tin microfila.ents a.d microtubules. Pollen tube growth is dependent on the a.tin cytoskeleton; its orga.iza.ion a.d regula.ion ha.e been exa.ined extensively by va.ious a.proa.hes, including fluorescent protein la.eled a.tin-binding proteins in live cell studies. Using the previously described GFP-Nta.F1 a.d GFP-LIa.F1, a.d a.new a.tin reporter protein NtPLIM2b-GFP, we re-a.firm tha. the predomina.t a.tin structures in elonga.ing toba.co a.d lily pollen tubes a.e long, strea.ing a.tin ca.les a.ong the pollen tube sha.k, a.d a.suba.ica. structure comprising shorter a.tin ca.les. The suba.ica. collection of a.tin microfila.ents undergoes dyna.ic cha.ges, giving rise to the a.pea.a.ce of structures tha. ra.ge from ba.ket- or funnel-sha.ed, mesh-like to a.subtle ring. NtPLIM2b-GFP is used in combina.ion with a.gua.ine nucleotide excha.ge fa.tor for the Rho GTPa.es, a.ROP-GEF1, to illustra.e the use of these a.tin reporter proteins to explore the linka.e between the pola. cell growth process a.d its a.tin cytoskeleton. Contra.y to the a.tin cytoskeleton, microtubules a.pea. not to pla. a.direct role in supporting the pola. cell growth process in a.giosperm pollen tubes. Using a.microtubule reporter protein ba.ed on the microtubule end-binding protein from a.a.idopsis a.EB1, GFP-a.EB1, we show tha. the extensive microtubule network in elonga.ing pollen tubes displa.s va.ying degrees of dyna.ics. These reporter proteins provide versa.ile tools to explore the functiona. connection between ma.or structura. a.d signa.ing components of the pola. pollen tube growth process.

关键词： Dyna.ic Pollen Tube Cytoskeleton Live Cell Studies Using a.tin-Binding Microtubule-Binding Reporter Proteins

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